REVIEWS

Salmonellae interplay with host cells

Andrea Haraga*, Maikke B. Ohlson‡ and Samuel I. Miller*‡§
Abstract | Salmonellae are important causes of enteric diseases in all vertebrates.
Characterization of the molecular mechanisms that underpin the interactions of
salmonellae with their animal hosts has advanced greatly over the past decade, mainly
through the study of Salmonella enterica serovar Typhimurium in tissue culture and animal
models of infection. Knowledge of these bacterial processes and host responses has
painted a dynamic and complex picture of the interaction between salmonellae and
animal cells. This Review focuses on the molecular mechanisms of these host–pathogen
interactions, in terms of their context, significance and future perspectives.

Pinocytosis
A nonspecific process by which
small volumes of extracellular
fluid are taken up by certain
eukaryotic cells owing to the
engulfment of fluid in small
membrane vesicles.
Tight junction
The connection between two
adjacent cells in a monolayer
that is formed by extracellular-
matrix and protein complexes;
impermeable to water and
other molecules.
Macropinocytosis
Used to refer to the
endo­cytosis of large volumes   layer and evade being killed by digestive enzymes, bile
of extracellular fluid and
particles by membrane ruffles.
*Department of Genome
Sciences, ‡Department of
Microbiology, §Department of
Medicine, University of
Washington, Seattle,
Washington 98195, USA.
Correspondence to S.I.M.
e‑mail: millersi@u.washington.
edu
doi:10.1038/nrmicro1788
Published online   tinal inflammatory responses, probably contributes
19 November 2007
Salmonellae are Gram-negative bacterial pathogens
that are capable of infecting a wide range of animals,
which can result in several manifestations of disease1.
The host specificities and the disease symptoms that are
caused by some of the thousands of Salmonella serovars
are listed in TABLE 1. Salmonellae are typically acquired
 
by the oral ingestion of contaminated food or water;
however, exposure to pet reptiles and amphibians, which
are often carriers of the bacteria, can also pose a risk for
salmonellosis in humans (FIG. 1). Although conditions that
 
increase the gastric pH reduce the infectious dose of the
bacterium, salmonellae have an adaptive acid-tolerance
response that might promote their survival in the low
pH milieu of the stomach2. After entering the small
intestine, salmonellae traverse the intestinal mucous
salts, secretory IgA, antimicrobial peptides and other
innate immune defences in order to gain access to the
underlying epithelium3–5. Salmonellae have the ability to
invade the non-phagocytic enterocytes of the intestinal
epithelium by bacterial-mediated endocytosis6. After
adherence to the apical surface of the cell, using vari-
ous fimbrial adhesins, the bacteria disrupt the epithelial
brush border and induce membrane ruffles that engulf
the organisms7,8. However, salmonellae preferentially
enter microfold (M) cells, which are specialized epithelial
cells that sample intestinal antigens through pinocytosis
and transport them to lymphoid cells in the underlying
Peyer’s patches9,10. Alternatively, they can translocate
through the intestinal epithelia after uptake by CD18-
expressing phagocytes11. Moreover, in vitro, salmonellae
are able to disrupt tight junctions, which seal the epithelial
cell layer and restrict the paracellular passage of ions,
water and immune cells12. This, in addition to intes-
to the induction of diarrhoea. The fact that there are
multiple mechanisms for crossing the intestinal barrier
indicates the importance of this strategy to the lifestyle
of salmonellae.
M-cell-mediated uptake also allows salmonellae
to interact with, and possibly enter, intestinal epithe-
lial cells through their basolateral surface. Epithelial
turnover and shedding then, in turn, could return
the bacteria to the lumen of the intestine, simply
requiring that they survive and, perhaps, replicate
intracellularly. Such mechanisms are possibly impor-
tant for intestinal colonization and even for acute
disease by salmonellae, as a great deal of data, from
both animal and cell culture models, indicate that the
specialized ability to invade and survive in epithe-
lial cells is essential for their pathogenesis 13–17. By
contrast, the importance of phagocytosis and traf-
ficking through CD18-positive cells is unknown and
might represent another mechanism, redundant to
M-cell-mediated entrance, for salmonellae to cross
the intestinal barrier without actively interacting with the
epithelial surface. It is interesting to speculate that this
method may be important for typhoid fever, in which so
few organisms can cause disease but induce only mini-
mal intestinal disturbance in the majority of cases. This
could allow Salmonella enterica serovar Typhi (S.  typhi)
to disseminate systemically in a relatively silent fashion,
as opposed to mechanisms that involve bacterial-medi-
ated endocytosis, which are associated with intestinal
inflammation and diarrhoea13,14.
Once the epithelial barrier has been breached,
Salmonella serotypes that are associated with systemic
illness can enter intestinal macrophages by inducing
macropinocytosis , sensing the phagosomal environ-
ment and activating various virulence mechanisms
in order to survive in the microbicidal environment
of the macrophage 18–25 . This promotes bacterial

nature reviews | microbiology volume 6 | january 2008 | 53

© 2008 Nature Publishing Group
REVIEWS
Table 1 | Examples of Salmonella enterica serovars; their hosts and diseases
Salmonella Host Disease and symptoms
enterica serovar specificity
Typhoid
Typhi; Human- Enteric fever: fever; abdominal pain; transient
Paratyphi restricted diarrhoea or constipation; and a salmon-
coloured maculopapular rash on the trunk
Non-typhoid
Typhimurium; Broad-range Gastroenteritis: abdominal pain; vomiting; and
Enteriditis inflammatory diarrhoea
replication and subsequent dissemination throughout
the reticulo­endothelial system (RES). Infection with
non‑typhoidal strains in healthy human adults, however,
is usually limited to the intestine, where the bacteria
induce an early inflammatory response that results in the
infiltration of polymorphonuclear leukocytes (PMNs)
into the intestinal lumen1,26. Production of the potent
PMN chemokine interleukin (IL)‑8, and, perhaps, other
similar molecules, by the infected epithelia is thought
to be required for this process27. The release of cyto-
toxic granules by PMNs, as well as the effects of various
bacterial molecules in stimulating innate immune
inflammatory responses and manipulating host-
cell processes, may then result in the destruction or
turnover of the intestinal mucosa, which contributes
to inflammatory diarrhoea.
Recent progress has been made in our understanding
of the molecular mechanisms by which salmonel-
lae interact with host cells and manipulate various host
responses that result in the induction of intestinal
inflammatory responses and systemic illness. Specifically,
the identification of bacterial and host proteins that are
involved in such processes as the invasion of epithelial
cells, stimulation and repression of signalling cascades,
sensing of the intracellular environment and establishment
of a niche for intracellular replication have contributed to
insights into the complex pathogenesis of this bacterium,
and will be discussed here in detail.
Reticuloendothelial system The two virulence-associated T3SSs
(RES). The meshwork of A specialized apparatus, named the type III secretion
connective tissue that contains
system (T3SS), is essential to salmonellae pathogen-
immune cells, such as
macrophages, and surrounds esis and the colonization of host tissues. The T3SS
tissues that are associated mediates the transfer of bacterial virulence proteins,
with the immune system, such known as effectors, from the bacterial cell into the
as the spleen and lymph host-cell cytoplasm28. Once inside the eukaryotic cell,
nodes. Immune cells in the RES
provide surveillance of antigens
these effectors can alter host cellular functions, such as
that the body encounters and cytoskeletal architecture, membrane trafficking, signal trans-
can be quickly recruited to duction and cytokine gene expression, in order to promote
sites of infection. bacterial survival and colonization. The T3SS is
evolutionarily related to the flagellar export system and
Pathogenicity island
A large region of genomic DNA is present in multiple Gram-negative animal and plant
that encodes genes that are pathogens. It comprises more than 20 proteins, some of
associated with virulence. A which form a supramolecular structure that is known as
pathogenicity island is typically the needle complex (NC) (FIG. 2). Electron micrographs of
transferred horizontally
between bacterial strains and
purified T3SSs revealed that the NC consists of a multi-
is often inserted into tRNA ring base that spans the inner and outer membranes of
genes within the genome. the bacterial envelope, an inner rod that joins the rings
54 | january 2008 | volume 6 www.nature.com/reviews/micro
© 2008 Nature Publishing Group
and a hollow needle29,30. Another set of proteins comprise
the translocon, which is thought to be involved in the
translocation of effectors by forming a translocation pore
in the host cellular membrane31. The effector proteins
have been shown to contain specific targeting signals
located in their amino termini that route them to the
T3SS32–35. In addition to the short secretion signal, many
effectors have a binding site for a specific chaperone,
the function of which is thought to be to stabilize and
target its cognate substrate to the translocation appara-
tus34,36,37. Most chaperones are specific for a single effec-
tor, but some can facilitate the secretion of more than one
effector protein38–41. An ATPase that is located in the base
of the NC not only drives the export process but also
facilitates the release of effectors from chaperones before
transport42. Although much of the T3SS is similar among
Gram-negative bacteria, the set of effector proteins is
unique to each species. For a complete list of Salmonella
spp. effectors, their host-cell functions and binding
partners, see Table 2.
Salmonellae encode two distinct virulence­­‑associated
T3SSs within Salmonella pathogenicity islands 1 and 2
(SPI1 and SPI2) that function at different times during
infection28. Whereas the SPI1-encoded T3SS is active
on contact with the host cell and translocates bacterial
proteins across the plasma membrane, the SPI2 T3SS is
expressed within the phagosome and translocates effectors
across the vacuolar membrane. The SPI1 system has been
shown to be required for invasion of non-phagocytic
cells, induction of intestinal inflammatory responses
and diarrhoea, as well as colonization of the intestine.
The SPI2 T3SS, by contrast, has an important role in
bacterial survival in macrophages and establishment of
systemic disease. Interestingly, Brown and colleagues43
have recently shown by RIVET (recombination-based
in vivo expression technology) that expression of the
SPI2 T3SS in mice might begin in the early stages of
Salmonella enterica serovar Typhimurium (S. typhimu-
rium) infection, before intestinal penetration. As there
is currently no evidence that this T3SS is also involved
in intestinal colonization, the authors speculated that its
early induction in the intestinal lumen might prepare
the bacterium for the inhospitable intracellular environ-
ment of the macrophage and ease the transition to the
later systemic phase of the disease. However, it is also
possible that early expression of this T3SS is required for
the optimal function of the invasion-associated T3SS, as
mutations in the SPI2 apparatus can cause a significant
reduction in the expression of several SPI1 T3SS genes
and impair the ability of the bacterium to invade epi-
thelial cells44,45. Conversely, there is mounting evidence
that some of the effectors of the SPI1 T3SS are expressed,
or persist within, host cells long after bacterial inter-
nalization and contribute to events that were previously
attributed exclusively to effectors of the SPI2 T3SS46–51.
Although it is unknown whether mutations in the SPI1
T3SS could cause pleiotropic effects, these findings indi-
cate that the two Salmonella T3SSs do not operate in an
independent manner, as previously thought, and pos-
sibly cooperate to facilitate the intracellular lifestyle of
the bacteria. This is particularly intriguing, because the

Salmonella spp.
M cell
Epithelial cell
Macrophage
T cell B cell
Gastroenteritis:
PMN influx
Enteric fever: dissemination
to lymph nodes, liver and spleen
Figure 1 | Biology of salmonellae infection. Orally ingested salmonellae
Nature Reviews |survive at the
Microbiology
low pH of the stomach and evade the multiple defences of the small intestine in order to
gain access to the epithelium. Salmonellae preferentially enter M cells, which transport
them to the lymphoid cells (T and B) in the underlying Peyer’s patches. Once across the
epithelium, Salmonella serotypes that are associated with systemic illness enter intestinal
macrophages and disseminate throughout the reticuloendothelial system. By contrast,
non-typhoidal Salmonella strains induce an early local inflammatory response, which
results in the infiltration of PMNs (polymorphonuclear leukocytes) into the intestinal
lumen and diarrhoea.
sequence information and genetic organization of the
two Salmonella T3SSs, as well as their phylogenetic
distribution among Salmonella species, indicate that
they were acquired independently, at different periods
and from different sources44,52–57. Therefore, it is likely
that selection pressure from animal environments led
to the cooperation between the two T3SSs, to opti-
mize colonization, invasion and intracellular survival
of salmonellae in animals.
The most extensively used animal model for investi-
gating the contributions of the two T3SSs to salmonellae
virulence is the highly sensitive natural-resistance-
associated macrophage protein 1 (Nramp1)‑null mouse,
which is susceptible to mortal infection if inoculated with
as few as ten bacteria58. Nramp1 is a macrophage-specific
ion exporter that removes ions from the Salmonella-
containing vacuole (SCV) and, thus, restricts bacterial
replication59. The Nramp1-null mouse model system
has been used successfully to identify many important
genes that are required for T3SS-associated virulence60,61.
However, it is too sensitive to detect phenotypes for
T3SS effectors that contribute to long-term persist-
ence, because these mice do not survive long enough
to study the proteins that are necessary for systemic
colonization. The resurgence of interest in the resistant
Nramp1-positive mouse model has enabled studies to
be performed that are more physiologically relevant
to long-term systemic infection49,62. These reports
confirmed that SPI2 T3SS effectors are important for
persistent infection and colonization, but also validated
nature reviews | microbiology volume 6 | january 2008 | 55
© 2008 Nature Publishing Group
REVIEWS
older data that had suggested a role for the SPI1 T3SS
during systemic disease17,63. This, combined with the
previously mentioned putative function of the SPI2 T3SS
prior to intestinal penetration, further demonstrates that
the dogmatic compartmentalization of the roles of the two
T3SSs is simplistic. In addition, the recent resurrection of
the streptomycin-treated mouse model of acute intestinal
infection with inflammation and diarrhoea might enable
dissection of the roles of the host and bacteria in acute
intestinal disease64. It is likely that, in the near future,
models of salmonellae-induced chronic intestinal disease
at the intestinal mucosal surface will be further devel-
oped and studied. In fact, our group has demonstrated
that the A/J mouse strain develops this type of disease
(L. Peiser, K. Smith and S.I.M., unpublished observa-
tions). These animal models should be useful in under-
standing the contributions of mucosal immune defences,
as well as bacterial factors such as T3SS effectors, to
chronic intestinal disease. However, these experimental
systems also have limitations, as specific disease outcomes
may be different in different hosts for broad host-range
salmonellae. A future challenge in understanding these
animal models will be to determine the effects of specific
effectors in both the model and natural hosts. In addi-
tion, the diversity and evolution of the effector content of
different Salmonella strains isolated from natural sources
might also reveal different functions or roles that cannot
be explored by these model systems. In this regard, many
S. typhimurium effectors are present in only a percentage
of strains, and in some serotypes, such as Typhi, are only
present as pseudogenes. Selection for the presence or
loss of effectors in specific animal populations or human
disease settings might, ultimately, be more informative of
the actual natural disease or colonization than the phe-
notypes of deletion mutants in model systems. Therefore,
the acceptance of the limitations of these animal mod-
els is important to prevent dogmatic viewpoints about
the roles of such mechanisms or specific T3SS effector
proteins. This is particularly relevant to the study of effec-
tor proteins with redundant functions, as the necessity
for their similar tasks during pathogenesis might not be
apparent in model systems. A complete picture will prob-
ably require the integration of animal model information
with real world information about effector content and
disease course in nature.
Bacterial-mediated endocytosis
The concerted function of at least five SPI1 T3SS effec-
tors is required for efficient invasion of cultured epithe-
lial cells, although optimal invasion in animal tissues
might be more complex and diverse (FIG. 3). SopE, SopE2
and SopB (also known as SigD) activate the host Rho
GTPases Cdc42, Rac1 and RhoG, which leads to actin
cytoskeletal reorganization, membrane ruffling and bac-
terial internalization by macropinocytosis65–70. However,
recent evidence suggests that only Rac1 and RhoG are
indispensable for the actin remodelling events that are
generated by Salmonella spp. during host-cell entry70.
SopE, SopE2 and SopB are all essential for the invasion of
epithelial cells, as an S. typhimurium mutant that is
defective in all three effectors cannot induce actin
REVIEWS
a modulating actin dynamics directly. SipA (also
Needle
Base
b
Base
Needle
Figure 2 | Structure of the Salmonella type III
Nature Reviews | Microbiology
secretion system (T3SS). a | Electron micrographs of a
purified Salmonella enterica serovar Typhimurium
(S. typhimurium) needle complex (NC). b | Electron
cryomicroscopy and surface images of the structures of
the base and NC of the S. typhimurium T3SS. Part a
reproduced, with permission, from Ref. 31 (2002)
Elsevier Science. Part b reproduced, with permission,
from Ref. 185  (2004) American Association for the
Advancement of Science.
rearrangements and, therefore, become intracellular69.
Whereas SopE and SopE2 are potent guanine nucleotide
exchange factors (GEFs) for all three GTPases, SopB
only stimulates Cdc42 and RhoG indirectly through its
phosphoinositide phosphatase activity65,67–70. Although
the mechanism of action of SopB is not yet understood,
Patel and colleagues70 have recently shown that activa-
tion of RhoG by SopB occurs by a cellular exchange
factor called SGEF. As SGEF has a phosphoinositide-
binding pleckstrin homology domain, it is plausible
that it is activated by the phosphoinositide fluxes that are
generated by the enzymatic action of SopB71. Activation
of Rho GTPases then results in the activation of the
Wiskott–Aldrich Syndrome protein (WASP) family
members N‑WASP and WAVE2, which leads to recruit-
ment of the actin-related protein‑2/3 (Arp2/3) complex
to sites of membrane ruffles and stimulation of actin
polymerization67,72–74.
There are two SPI1 T3SS effectors that promote
bacterial internalization by binding to actin and
56 | january 2008 | volume 6 www.nature.com/reviews/micro
© 2008 Nature Publishing Group
called SspA) helps to initiate actin polymerization
at the site of S. typhimurium entry by decreasing the
critical concentration and increasing the stability
of actin filaments, whereas SipC (also called SspC),
an effector that also functions as a translocon and is
inserted in the host cell’s plasma membrane with a
cytoplasmic domain that may bind to intermediate
filaments, can nucleate and bundle actin 75–77. In
addition, SipA can enhance the activity of SipC
independently of any host cellular protein, which
indicates that there might be a unique collaboration
between the two effectors78. Although SipA and SipC
might act in concert with SopE, SopE2 and SopB to
mediate the cytoskeletal changes that are required
for the formation of membrane ruffles, they can-
not induce membrane ruffling and invasion by
themselves 69,76,77,79. Instead, they seem to facilitate
efficient bacterial uptake by directing the spatial
localization of actin foci beneath the invading bac-
teria and, perhaps, preventing disassembly of the
S. typhimurium-induced actin structures.
Intestinal inflammatory responses
The stimulation of Cdc42 by SopE, SopE2 and SopB
also triggers several mitogen-activated protein kinase
(MAPK) pathways, including the Erk, Jnk and p38 path-
ways, which results in the activation of the transcription
factors activator protein‑1 (AP‑1) and nuclear factor-κB
(NF-κB)16,70,80 (FIG. 3). These transcription factors then
direct the production of pro-inflammatory cytokines,
such as IL‑8, which stimulate PMN transmigration and
the inflammatory response leading to diarrhoea. Boyle
and colleagues81 have recently reported that disrup-
tion of tight junction structure and function — one
aspect of S. typhimurium-induced enteritis that is
likely to be important — by the effectors SopB, SopE,
SopE2 and SipA also occurs by activation of Rho fam-
ily GTPases. In addition, SopB also promotes intestinal
disease by increasing the intracellular concentration of
d‑myo-inositol 1,4,5,6-tetrakisphosphate, a compound
that stimulates cellular chloride secretion and fluid
flux69,82. SipB (also called SspB), an effector protein that
is also part of the SPI1 T3SS translocation machinery,
adds to the inflammatory response by increasing pro-
duction of the pro-inflammatory cytokines IL‑1β and
IL‑18 through binding and activating caspase‑1 (Ref. 83).
Caspase‑1 activation can also occur by the recognition of
flagellin by the Ipaf cytoplasmic signalling pathway84,85.
It is plausible that this is the result of the accidental
translocation of flagellin into the host cell’s cytoplasm
by the SPI1 T3SS owing to the conserved evolution of
the flagellar and the SPI1-encoded secretion systems.
The observation that caspase‑1-deficient mice are more
susceptible to orally administered S. typhimurium than
wild-type mice indicates that SPI1 T3SS-mediated cas-
pase‑1 activation could be essential to diarrhoeal disease
and intestinal survival86,87. SopA and SopD also contrib-
ute to enteritis in calves13,88,89. Recent evidence indicates
that SopA has a HECT (homologous to E6-AP carboxyl
terminus)-like E3 ubiquitin ligase activity, which is
 REVIEWS

Table 2 | Effectors of the Salmonella SPI1- and SPI2-encoded type III secretion systems (T3SSs)
Effector Cellular function Host-cell target References
SPI1 T3SS
AvrA Inhibits nuclear factor (NF)‑κB signalling and interleukin (IL)‑8 Unknown 99,186
production; also prevents ubiquitination of β‑catenin
SipA or SspA Decreases the critical concentration of G‑actin and increases the F-actin; T‑plastin 76, 81,187,188
stability of F‑actin; also induces PMN transepithelial migration
and disrupts tight junctions
SipB or SspB* Binds and activates caspase‑1 and induces autophagy in Caspase‑1; cholesterol 83,189,190
macrophages
SipC or SspC* Nucleates and bundles actin F-actin; cytokeratin‑8 and 75,77,191
cytokeratin-18
SopA Stimulates PMN transmigration by HECT-like E3 ubiquitin ligase Unknown 89,90
activity
SopB or SigD Activates Cdc42, RhoG, AktA and chloride secretion through its Unknown 69,70,81,82,192
inositol phosphatase activity and disrupts tight junctions
SopD Stimulates fluid accumulation in bovine ligated ileal loops and Unknown 13,82,91
contributes to diarrhoea in calves and systemic disease in mice
SopE Activates Cdc42, Rac1 and RhoG by its GEF activity and disrupts Cdc42, Rac1 and Rab5 65,81,193
tight junctions
SopE2 Activates Cdc42, Rac1 and RhoG by its GEF activity and disrupts Cdc42 and Rac1 67,68,81
tight junctions
SptP Inhibits Cdc42 and Rac1 by its GAP activity and MAPK signalling Rac1 92,94,95
and IL‑8 secretion through its tyrosine phosphatase activity
SPI2 T3SS
GogB Unknown Unknown 194
PipB Unknown Unknown 157
PipB2 Contributes to Sif formation Kinesin‑1 157,165,170
SifA Induces Sif formation, maintains integrity of the SCV and SKIP and Rab7 154,164,166, 181,195
downregulates kinesin recruitment to the SCV
SifB Unknown Unknown 156
SopD2 Contributes to Sif formation Unknown 91
SpiC* Interferes with endosomal trafficking Hook3 150,151–153
SpvB‡ Actin-specific ADP-ribosyltransferase and downregulates Sif Actin 160, 175,176,196
formation
SseF Contributes to Sif formation and microtubule bundling Unknown 162,169
SseG Contributes to Sif formation and microtubule bundling Unknown 162,169
SseI or SrfH Contributes to host-cell dissemination Filamin and TRIP6 160,197
SseJ Maintains integrity of the SCV and has deacylase activity Unknown 155,180
SseK1 Unknown Unknown 198
SseK2 Unknown Unknown 198
SseL Deubiquitinase Ubiquitin 199
SspH2 Inhibits the rate of actin polymerization and contributes to Filamin and profilin 96,160
virulence in calves
SteA Unknown Unknown 200
SteB Unknown Unknown 200
SteC Unknown Unknown 200
SPI1 and SPI2 T3SS
SlrP Contributes to virulence in calves Unknown 201
SspH1 Inhibits NF‑κB signalling and IL‑8 secretion, contributes to PKN1 95–98
virulence in calves and has E3 ubiquitin ligase activity
*Also a component of the secretion apparatus. ‡Has not been definitively shown to be an SPI2 T3SS effector. GAP, GTPase-activating protein; GEF, guanine
nucleotide exchange factor; HECT, homologous to E6-AP carboxyl terminus; MAPK, mitogen-activated protein kinase; PMN, polymorphonuclear leukocyte; SCV,
Salmonella-containing vacuole; Sif, Salmonella-induced filament; SPI, Salmonella pathogenicity island.

nature reviews | microbiology volume 6 | january 2008 | 57

© 2008 Nature Publishing Group
REVIEWS

a multifunctional protein that is also involved in the

Membrane reversal of the pro-inflammatory signalling cascade
Tight
ruffle
junction that is associated with invasion94. It has tyrosine phos-
Salmonella phatase activity, which has a role in downregulation
spp.
of the S. typhimurium-induced activation of the MAPK
Actin
Erk. In addition, S. typhimurium can downregulate IL‑8
production after invasion of intestinal epithelial cells,
SopE SipA SopA and SptP and SspH1 participate in this process95. SspH1
Epithelial
cell
SopE2 SipC SptP is a leucine-rich-repeat protein that is localized to the
SopB
SopB Rho GTPases mammalian nucleus and inhibits NF‑κB-dependent
gene expression95,96. It has also been shown to have
Rho GTPases MAPK CI– MAPK SspH1
AvrA ubiquitin ligase activity97. As it interacts with a host
NF-κB
CI– NF-κB serine/threonine protein kinase called PKN1, which if
AP-1 activated also inhibits NF‑κB-dependent gene expres-
sion, SspH1 could interfere with inflammatory signal-
ling by binding to and activating PKN1 (Ref. 98). In fact,
IL-1β
IL-8
IL-18 PKN1 is activated in epithelial cells during infection
with S. typhimurium. However, as this also occurs in
Macrophage the absence of SspH1, it is possible that SspH1 is not
SipB required for this process or that there are additional
PMN
effectors or bacterial mechanisms that are capable of
activating PKN1. Another SPI1 T3SS effector, AvrA,
Figure 3 | SPI1 T3SS-induced changes in host cells. On contact with the epithelial has been reported to inhibit NF‑κB activity and pro-
Nature Reviews | Microbiology
cell, salmonellae assemble the Salmonella pathogenicity island 1(SPI1)-encoded type III inflammatory cytokine secretion99. However, unlike
secretion system (T3SS) and translocate effectors (yellow spheres) into the eukaryotic its Yersinia spp. homologue, YopJ, which binds and
cytoplasm. Effectors, such as SopE, SopE2 and SopB, then activate host Rho GTPases, acetylates MAPK kinases and the inhibitor of NF-κB
which results in the rearrangement of the actin cytoskeleton into membrane ruffles, kinase (IKK), thereby preventing their phosphorylation
induction of mitogen-activated protein kinase (MAPK) pathways and destabilization of
and activation, AvrA has not been shown to interact
tight junctions. Changes in the actin cytoskeleton, which are further modulated by the
actin-binding proteins SipA and SipC, lead to bacterial uptake. MAPK signalling with or affect the activity of any host proteins that are
activates the transcription factors activator protein‑1 (AP‑1) and nuclear factor-κB involved in the NF‑κB pathway100,101.
(NF‑κB), which turn on production of the pro-inflammatory polymorphonuclear The fact that salmonellae use multiple effectors to
leukocyte (PMN) chemokine interleukin (IL)‑8. SipB induces caspase‑1 activation in dampen the host’s inflammatory response indicates
macrophages, with the release of IL‑1β and IL‑18, so augmenting the inflammatory that this function is important for their pathogenesis,
response. In addition, SopB stimulates Cl– secretion by its inositol phosphatase activity. perhaps by contributing to colonization of the intestinal
The destabilization of tight junctions allows the transmigration of PMNs from the tract over prolonged periods of time. Furthermore, it is
basolateral to the apical surface, paracellular fluid leakage and access of bacteria to the interesting to speculate that the evolutionary pressure
basolateral surface. However, the transmigration of PMNs also occurs in the absence of on the host–pathogen interaction for non-typhoidal
tight-junction disruption and is further promoted by SopA. The actin cytoskeleton is
salmonellae is for asymptomatic intestinal colonization,
restored and MAPK signalling is turned off by the enzymatic activities of SptP. This also
results in the down-modulation of inflammatory responses, to which SspH1 and AvrA whereas for typhoidal strains, which have poor intesti-
also contribute by inhibiting activation of NF‑κB. Figure reproduced from an original nal colonization, the selective pressure is for replication
kindly provided by A. Haraga, University of Washington, USA. within professional phagocytes, a condition that is also
less inflammatory than extracellular growth. The bacte-
ria might induce a mild state of inflammatory response
involved in the S. typhimurium-induced transepithelial and their goal might be to evolve towards parasitism or
migration of PMNs90. However, how SopD, an effector that commensalism. In fact, the long period of intestinal colo-
is also expressed under SPI2 T3SS-inducing conditions nization greatly exceeds that of the acute disease phase for
and continues to contribute to later stages of infection, non-typhoidal strains102. Also, typhoidal disease in the
exerts its effects remains unknown33,91. In summary, all absence of antibiotic therapy is often characterized by
of these mechanisms strongly implicate the SPI1 T3SS in periods of few or no symptoms, followed by a relapse that
the induction of gastroenteritis and intestinal disease. is more of a chronic, prolonged febrile illness, in which
intracellular replication is the predominant bacterial
Reversal of cytoskeletal and signalling responses lifestyle1. Thus, it is likely that a dynamic induction of
Interestingly, shortly after bacterial invasion the actin inflammatory responses, followed by their dampening, is
Transepithelial migration
cytoskeleton regains its normal architecture7. SptP has important to the diseases caused by salmonellae.
The movement of cells, such as
neutrophils and invading been shown to participate in this reversal process by
bacteria, from the basolateral its GTPase-activating protein activity on Cdc42 and The SCV as a niche for replication
(bottom) to the apical (top) Rac1 (Ref. 92) (FIG. 3). The opposing activities of SopE Salmonellae are capable of infecting and replicating in
surface, or the reverse, of an or SopE2 and SptP are mediated by the temporal regula- many cell types, but are thought to primarily replicate
epithelial cell layer. Migration
can also occur between two
tion of these proteins93. SopE has a shorter half-life in in macrophages, as they are found in the lymphatic tis-
adjacent cells through tight eukaryotic cells than SptP owing to its ubiquitination sues and organs of the RES during systemic infection1,103.
junctions. and rapid proteasome-dependent degradation. SptP is Salmonella strains that are defective for macrophage

58 | january 2008 | volume 6 www.nature.com/reviews/micro

© 2008 Nature Publishing Group
 REVIEWS
Salmonella spp.

intracellularly from hours to days, making it a unique

SspH2 SifA
SpvB PipB2 s macrophages in which the lysosomal compartments
Spacious Ssel tor
phagosome le mo
u
tub
M icro Sif
SCV
Actin SseJ
Epithelial cell
Microtubules
Nucleus
SseF
SseG
Golgi Secretory vesicles
Figure 4 | Formation of the SCV and induction of the SPI2 Nature
T3SSReviews Microbiology
within| the host cell.
Shortly after internalization by macropinocytosis, salmonellae are enclosed in a
spacious phagosome that is formed by membrane ruffles. Later, the phagosome fuses
with lysosomes, acidifies and shrinks to become adherent around the bacterium. This is
called the Salmonella-containing vacuole (SCV), which contains the endocytic marker
lysosomal associated membrane protein 1 (LAMP‑1; purple). The Salmonella
pathogenicity island 2 (SPI2) T3SS (type III secretion system) is induced within the SCV
and translocates effector proteins (yellow spheres) across the phagosomal membrane
several hours after phagocytosis. The SPI2 T3SS effectors SifA and PipB2 contribute to
Salmonella-induced filament (Sif) formation along microtubules (green) and regulate
microtubule-motor (yellow star shape) accumulation on the Sif and the SCV. SseJ is a
deacylase that is active on the phagosome membrane. SseF and SseG cause
microtubule bundling adjacent to the SCV and direct Golgi-derived vesicle traffic
toward the SCV. Actin accumulates around the SCV in a SPI2 T3SS-dependent manner,
in which SspH2, SpvB and SseI are thought to have a role.
replication are avirulent in mouse models of infection,
which underscores the importance of bacterial survival
and replication in macrophages to disease104. In addi-
tion, the observation that many attenuated strains of
salmonellae are auxotrophic, particularly for purines,
pyrimidines, amino acids and other metabolites that
are required for their replication but that are not readily
available within mammalian cells, supports the concept
that intracellular replication is essential to salmonellae
virulence105. Unlike non-phagocytic cells, salmonellae
can enter macrophages by several endocytic processes,
including SPI1 and non-SPI1 T3SS-induced macropino-
cytosis20. Following SPI1 T3SS-induced macropinocyto-
sis, a few SPI1 T3SS effectors, such as SipA, SopB, SopD
Auxotrophic
An organism that cannot
and SopE2, persist within the cell and have recently been
synthesize certain organic implicated in contributing to the intracellular stages of
compounds, such as amino the infection process47,48,50,51,91,106. Once intracellular,
acids, that are necessary for its salmonellae remain inside a vacuolar compartment,
metabolism. For growth,
which has been named the spacious phagosome (SP)20
auxotrophic organisms must
be able to take up the lacking (FIG. 4). The SP shrinks over minutes to hours to form
compound from the an adherent membrane around one or more bacteria,
surrounding environment. which is then referred to as the SCV. The SCV can persist
phagosome with respect to the normal progression of
phagolysosomal maturation and recycling. Although
there has been controversy within the field, several
reports have shown that salmonellae can survive within
have fused with the SCV107–110. Consequently, the avoid-
ance of phagolysosomal fusion is unlikely to be a major
pathogenic strategy of salmonellae. Studies in various
cell types have also demonstrated that the vacuole acidi-
fies; however, depending on the mechanism of host‑cell
entry, vacuolar acidification may be delayed in both
macrophages and epithelial cells19,110–112. The SCV has
been shown to interact transiently with the early endo-
cytic pathway and quickly gain and lose early endocytic
markers, such as EEA1 (early endosomal antigen 1), TfR
(transferrin receptor) and the early endocytic traffick-
ing GTPases Rab5 and Rab11 (Refs 113,114). Several late
endosomal markers are commonly associated with the
SCV at later time points, including the GTPase Rab7,
LAMP1 (lysosomal associated membrane protein 1),
LAMP2, LAMP3 and the vacuolar ATPase110,113,115,116.
There is conflicting data concerning the occurrence of
M6PR (mannose‑6-phosphate receptor), LBPA (lysobi-
sphosphatidic acid) and the lysosomal hydrolase cathep­
sin D on the SCV115,117–120. Furthermore, cholesterol has
been reported to accumulate on the SCV118,121. The pres-
ence or absence of different markers on the persistent
SCV may simply indicate that they are variably detected
rather than reflect whether or not the SCV has matured
through a normal endocytic pathway. As it has been
shown that the SCV can fuse with lysosomes and acidify,
the ability of salmonellae to survive exposure to lyso-
somal contents reveals that resistance to antimicrobial
peptides, nitric oxide and oxidative killing are important
to its survival within macrophages and to virulence122–129.
This is supported by the observations that Salmonella spp.
mutants that are sensitive to these chemicals are attenu-
ated for mouse virulence, whereas mice that are deficient
for the production of these compounds have increased
susceptibility to Salmonella spp.23,104,129–131
Sensing and response to the vacuole
Salmonellae sense the acidic environment of the SCV,
resulting in the induction of various regulatory systems
that promote intracellular survival, for example, by
surface remodelling of the protein, carbohydrate and
membrane components of the bacterial envelope19. Such
regulatory systems include OmpR/EnvZ, PhoP/PhoQ,
RpoS/RpoE, PmrA/PmrB, Cya/Cyp and cyclic diGMP,
all of which confer resistance to antimicrobial peptides
and oxidative stress18,124,127,129,132–135. The phagosomal
environment is acidic, with a pH range of <5 to 5.5, has
a concentration of magnesium and calcium in the 1 mM
range and contains antimicrobial peptides and oxygen
and nitrogen radicals that can damage the bacterial
cell19,111,112. Various studies indicate that both pH and
antimicrobial peptides are important signatures of the
phagosomal environment and such conditions activate
many of the regulators that are implicated in salmonellae
virulence24,25,125,136. It is likely that several sensory systems

nature reviews | microbiology volume 6 | january 2008 | 59

© 2008 Nature Publishing Group
REVIEWS
respond to the phagosome environment and cooperate
to orchestrate the complex cascade of events that are nec-
essary to alter the bacterial surface and promote intracel-
lular survival137. The best characterized of these is the
PhoQ sensor, which promotes resistance to antimicro-
bial peptides18,25. PhoQ contains a novel acidic domain
that is bridged to the inner membrane by interactions
with metal ions and binds to and initiates responses to
antimicrobial peptides138,139. It also responds to pH by
structural changes that are determined by regions of the
protein separate from the metal ion bridges involved in
antimicrobial peptide sensing136.
After phagocytosis, salmonellae undergo extensive
bacterial surface remodelling, as has been shown for the
lipid A component of lipopolysaccharide (LPS) during
growth within macrophages137,140. Bacterial molecules
that the host can recognize as indicators of infection,
such as the SPI1 T3SS and flagellin, are repressed and
the LPS structure is altered85,141. Some of the specific sur-
face modifications include: decreasing the length of the
O antigen, which is the repeating carbohydrate polymer
of LPS; alterations to the number of acyl chains in the
structure of the lipid A component of LPS; and changes
in the protein content of the outer membrane, the inner
membrane and the peptidoglycan layer142–145. Synthesis
of enzymes that allow the bacteria to cope with oxida-
tive and nitrogenous radicals also occurs146. Microarray
studies have shown that up to 919 S. typhimurium
genes are differentially regulated in response to the
phagosomal environment, demonstrating that dra-
matic transcriptional and post-translational changes
probably occur when salmonellae make the transition
from a nutrient-rich extracellular environment to the
intracellular environment147.
Protein delivery across the vacuolar membrane
As a result of sensing the phagosomal environment,
the expression and assembly of the SPI2-encoded T3SS
is induced134,148. Although the function of this T3SS in
pathogenesis remains poorly defined, it has been shown
to be essential for virulence in the mouse model of infec-
tion22,149. As mutants of the SPI2 T3SS cannot replicate
in tissue-culture cells and animal models, it is likely
that the role of this T3SS during disease is to promote
intracell­ular replication within the intestine during the
acute phase of the infection and in other organs during
the chronic state. A reasonable hypothesis for the main
function of the SPI2 T3SS is that it promotes intracellular
replication by altering host vesicular trafficking, so that
useful metabolic molecules, such as amino acids and lip-
ids, are routed to the SCV and the vesicular compartment
membrane is expanded. To date, at least 20 Salmonella
spp. effector proteins are known to be translocated by
the SPI2 T3SS across the phagosomal membrane into the
eukaryotic-cell cytoplasm; however, their specific roles
in promoting intracellular replication or modifying
vesicular movement are not yet understood (for a list
of SPI2 T3SS effectors, their functions and binding part-
ners, see TABLE 2). In addition, no individual translocated
effector has definitively been shown to alter vesicular
trafficking. Although it has been reported that SpiC alters
60 | january 2008 | volume 6 www.nature.com/reviews/micro
© 2008 Nature Publishing Group
endosome–endosome fusion in vitro and binds to pro-
teins that are implicated in vesicular trafficking, its role
as a T3SS effector protein remains controversial150,151. As
it has not been universally observed to be translocated
into eukaryotic cells and it is required for the transloca-
tion of several, if not all, SPI2 T3SS effectors, as well as
the surface expression of translocon proteins, it is more
likely that SpiC is part of the SPI2 secretion apparatus
and is not an effector152,153. The most important trans-
located effectors, by virtue of causing virulence defects
in mice when mutated, are SifA, SseJ, SseF, SseG, SopD2
and PipB2 (Refs 91,154–158) . The observation that
S. typhimurium that lacks any single SPI2 T3SS effector
protein cannot cause the same virulence attenuation in
mice as a mutant strain that lacks the entire SPI2 T3SS
suggests that many effectors function cooperatively to
exert their effects on the host cell. Furthermore, the
deletion or mutation of many effector genes has no
virulence phenotype, which implies that their functions
might be redundant. Early studies of SPI2 T3SS effectors,
which primarily focused on determining their subcel-
lular localization in mammalian cells, revealed that
they might have specific targeting sequences that direct
localization to endosomal compartments, the Golgi
apparatus, the actin cytoskeleton and the microtubule
network33,155–157,159–162. This indicates that components
of these host-cell structures might be the intracellular
targets of the SPI2 T3SS.
Salmonella-induced tubular endosomes
In epithelial cells and interferon‑γ-primed macrophages,
the SPI2 T3SS induces the formation of long filamen-
tous membrane structures — Salmonella-induced fila-
ments (Sifs) — that originate from the SCV and extend
throughout the cell157,163 (FIG. 4). These structures are
LAMP1-positive and have a similar membrane composi-
tion to the SCV118. Sif formation requires microtubules,
but not the actin cytoskeleton, although these filaments
can be decorated with actin159. Possible mechanisms that
contribute to Sif formation include repetitive initiation
of vesicular budding from the SCV, in which fission
events are incomplete, or continuous fusion of endocytic
vesicles with the SCV, which would result in endosomal
tubulation or elongation of the SCV. Regardless of the
mechanism of Sif formation, the filaments may func-
tion to increase the size of the SCV to accommodate
bacterial replication during systemic infection. Sif
formation is dependent on the SPI2 T3SS effector SifA,
and to a lesser extent on SseF, SseG, SopD2 and PipB2
(Refs 91,164,165). However, it is a dynamic phenotype
that may also be modulated by other effectors. SifA was
recently shown to bind to the host cell protein SKIP (SifA
and kinesin interacting protein)166. The same authors
also found that, if SKIP was depleted by RNA interfer-
ence in S. typhimurium-infected cells, the cells failed to
form Sifs, suggesting that Sif formation requires SKIP.
In addition, Alto and colleagues167 identified SifA as a
member of the WxxxE (tryptophan (W)-variable (x)-x-
x-glutamate) family of bacterial effectors that function as
mimics of GTPases. As SifA contains a carboxy-terminal
Caax (cysteine (C)-aliphatic residue (a)-a-x) motif that

Prenylated
The post-translational addition
of lipid chains, such as farnesyl
or geranylgeranyl, to cysteine
residues in proteins that
contain a prenylation motif
called a CaaX box. This
process facilitates membrane
localization and/or protein–
protein interactions.
nature reviews | microbiology volume 6 | january 2008 | 61
is prenylated and S‑acylated by host-cell enzymes, similar
to GTPases, it is possible that SifA uses a GTPase-type
mechanism for membrane localization and, perhaps, Sif
formation168. Furthermore, the region of SKIP that inter-
acts with SifA is a pleckstrin homology domain, which
is commonly found in proteins that bind to signalling
lipids and other regulatory molecules, including those
that modulate Rho GTPases166.
The roles of PipB2, SopD2, SseF and SseG in Sif for-
mation are not entirely clear. S. typhimurium strains that
lack any of these effectors do not induce Sifs as efficiently
as wild-type S. typhimurium91,165,169. Instead, infection of
cultured cells with these mutants tends to lead to the for-
mation of ‘ pseudo-Sifs’, which extend from the SCV and
co-localize with effectors, but do not contain LAMP1.
This suggests that in the absence of these proteins the
ability to form Sifs — which is defined as being entirely
LAMP1-positive — is impaired and that induction of
Sifs involves multiple steps that are orchestrated by dif-
ferent effectors. Transient expression of PipB2 in mam-
malian cells induces the movement of LAMP1-positive
compartments to the cell periphery, which is probably
the result of its interaction with the plus-end-directed
microtubule motor kinesin165,170. This activity might
contribute to the outward extension of the SCV, which
would promote Sif formation. SopD2, which has homo­
logy to the SPI1 T3SS translocated effector SopD, has
also been shown to cooperate with SifA to induce Sifs,
but by an as yet unidentified mechanism33,91. SseF and
SseG promote the aggregation of endosomal vesicles
and recruit Golgi-derived exocytic vesicles to the SCV,
which suggests that salmonellae are able to usurp both
endocytic and exocytic cellular transport processes171–174.
However, it is not known how these activities directly
influence Sif formation.
The deletion of SseJ and SpvB can cause an increase
in Sif formation at later time points during the infection
of cultured cells, which suggests that these proteins have
Sif downregulatory functions175. Furthermore, these
effectors have virulence defects in the Nramp1-null ani-
mal model, which indicates that their activities contrib-
ute to systemic infection156,176. Although SpvB has not
been formally demonstrated to be an SPI2 T3SS effector,
some studies suggest that the SPI2 T3SS is required for
its activity in infected cells177,178. SpvB is an actin-specific
ADP-ribosyltransferase that promotes actin depolym-
erization176. SseJ has homology to glycerophospholipid
cholesterol acyl transferase enzymes, which can remove
acyl chains from phospholipids and transfer them to
cholesterol in a two-step deacylase-acyltranferase
reaction32,179. In fact, purified SseJ has deacylase activ-
ity in vitro, which has been shown to be required for
its virulence in mice180. It has been proposed that SseJ
and SifA have complementary roles in maintaining the
integrity of the SCV membrane, because sifA-mutant
S. typhimurium tends to lose the SCV membrane but
does not do so if SseJ is also lacking. This suggests that,
in the absence of SifA, SseJ could cause damage to the
SCV by its enzymatic activity. In addition, our labora-
tory has obtained evidence that SseJ has phospholipase
activity in vivo that might be localized to the endosomal
© 2008 Nature Publishing Group
REVIEWS
membrane, which has led us to propose that its most
likely function during S. typhimurium infection is to
cleave phospholipids from the SCV membrane (Megha,
M.B.O. and S.I.M., unpublished observations). The
cleavage of acyl chains from phospholipids could then
promote curvature of the membrane or produce discrete
lipid environments that serve as platforms for promot-
ing vesicle fusion and binding scaffolding proteins and
such activity could modulate Sif formation. However,
how these functions, and Sif formation in general, are
important for Salmonella-induced disease is currently
not understood. It is plausible that Sifs increase the size
of the SCV in a specific and directional fashion that
promotes bacterial replication and/or redirect nutri-
ent-rich organelles to the SCV. In this regard, the SPI2
T3SS could help salmonellae to replicate inside the
phagosome and gain important nutrients for growth,
but yet avoid some of the host-defence mechanisms and
inflammatory responses that would result from their
release into the cytoplasm.
Microtubule motors and movement of the SCV
As Sif formation requires microtubules — which have
been observed to accumulate and bundle around the
SCV — there has been an increased interest in under-
standing how microtubules and microtubule motors
contribute to the intracellular life cycle of salmonellae
and whether targeting vesicular trafficking along micro-
tubule networks in infected cells is indeed one of their
pathogenic strategies162 (FIG. 4). SifA has been reported to
downregulate the recruitment of the plus-end directed
microtubule motor kinesin to the SCV, which is medi-
ated by PipB2 (Refs 166,170). It does so by interacting
with SKIP, which, if in a complex with SifA, displaces
kinesin from the SCV166. This interference is thought to
be crucial for maintaining the integrity of the SCV. The
inhibitory activity of SifA on kinesin seems to be domi-
nant over the kinesin-recruiting activity of PipB2, which
is suggestive of a potential coordinate or temporal regu-
lation between the two effectors170. In addition, SseF and
SseG have been found to co-localize with microtubules
and cause microtubule bundling in S. typhimurium-
infected cells. In particular, SseF has been implicated
in recruiting the minus-end directed motor dynein to
the SCV162,173. Therefore, it would appear that the SCV
accumulates dynein, but not kinesin. However, as SifA
inhibits the interaction between Rab7 and its effector
RILP, a protein that is associated with the dynein motor
complex, it is plausible that SifA also prevents dynein
accumulation on the SCV181,182. Although these results
paint a paradoxical picture of the need to promote or
inhibit microtubule-motor accumulation on the SCV,
salmonellae might employ a carefully choreographed
combination of both the recruitment and inhibition
of both types of microtubule motors. For example, at
specific times during infection, the recruitment of
dynein could promote minus-end-directed movement
of the SCV to stabilize it near the nucleus or the Golgi
apparatus, where the SCV would have increased access
to trafficking vesicles that contain nutrients and mem-
brane, whereas at other times the recruitment of kinesin
REVIEWS
to the SCV could induce plus-end-directed movement
towards the cell periphery to promote the enlargement
of the SCV and/or elongation of Sifs.
Recent studies have also shown that the SCV
in infected cultured epithelial cells is localized in a
perinuclear position and that effectors from both the
SPI1- and the SPI2-encoded T3SSs are involved in this
process50,161,166,170,173,181. In particular, SifA, SseF, SseG
and SipA seem to control this phenomenon, because
the SCV containing S. typhimurium strains lacking any
of these effectors does not localize near the nucleus. As
the disruption of perinuclear localization of the SCV is
correlated with decreased bacterial replication, it has
been proposed that salmonellae require juxtanuclear
positioning for optimal growth. In fact, Salcedo and
Holden161 have shown that SCVs in close proximity
to the nucleus contain more bacteria on average than
SCVs further away from the nucleus. However, it
has not been directly demonstrated that non-nuclear
localization of the SCV, per se, is inhibitory to bacterial
replication. Rather, it seems more likely that the lack
of these effectors or disruption of vesicle trafficking
along the microtubule network correlates with defective
replication and that non-nuclear SCV positioning is a
by-product of this more direct effect. Furthermore, the
positioning of the SCV might be different in polarized
epithelial cells, in which movement of the vacuole in a
directional fashion, such as during transcytosis, could
be more physiologically relevant. As discussed above, it
is possible that positioning the SCV in close proximity
to the Golgi, or other organelles, has some advantages
for salmonellae in terms of acquiring nutrients, inhibit-
ing immune responses, prolonging the maturation of
intestinal epithelial cells and allowing more intracellular
time before epithelial shedding into the intestinal lumen.
All of these effects could then contribute to intestinal
colonization or the intracellular growth of salmonellae.
Actin polymerization around the SCV
SPI2 T3SS-dependent actin condensation around the
SCV has been observed, which suggests that this T3SS
is also involved in cytoskeletal modifications160,183 (FIG. 4).
In addition, the treatment of S. typhimurium-infected
cells with inhibitors of actin polymerization results in
decreased bacterial replication, which indicates that the
manipulation of the actin cytoskeleton is important for
bacterial replication183. Although the effectors that are
responsible for the recruitment of actin to the SCV have
not been identified, several have been shown to interact
with actin-binding proteins or to directly manipulate
actin polymerization. However, it is also possible that
SCV-associated actin polymerization is not mediated
by effectors, but is due to the insertion of the T3SS
translocon into the SCV membrane, which has also
been observed with the similar Yersinia spp. T3SSs184.
Indeed, most SPI2 T3SS effectors that have been shown to
interact with components of the actin cytoskeleton have
actin-inhibitory activities, which suggests that the most
likely goal of salmonellae is to reduce or reverse vacuole-
associated actin polymerization (VAP). For example,
SpvB has been shown to be involved in the disassembly
62 | january 2008 | volume 6 www.nature.com/reviews/micro
© 2008 Nature Publishing Group
of VAP, presumably by ADP-ribosylating actin and
promoting actin depolymerization160,176. SspH2 has also
been reported to inhibit the rate of actin polymerization
in vitro160. In addition, SspH2 can interact with newly
polymerized actin, probably by binding to the actin-
binding proteins filamin and profilin, through its amino
and carboxy termini, respectively. Another effector, SseI
(also called SrfH), shares a high degree of identity with
the amino terminus of SspH2 and has been shown to
bind to polymerized actin, probably through filamin.
Although transiently expressed SspH2 and SseI are
associated with VAP during S. typhimurium infection,
they are not required for the formation of this structure.
Thus, unlike the clearly defined and temporally control-
led roles of the SPI1 T3SS effectors in manipulating the
actin cytoskeleton during invasion of epithelial cells, the
biological significance of the SPI2 T3SS-mediated actin
recruitment to the SCV, and the interaction of the effec-
tors of this T3SS with the actin network, remains to be
understood. It might be important for translocon stabil-
ity and translocation, thereby allowing the full effector
complement to be delivered consistently, and it could
explain the importance of this process to intracellular
replication183. Subsequently, as T3SS effectors seem to
decrease or remodel actin polymerization, there might
be a period in which actin removal is important, for
example, to decrease inflammatory responses or allow
fusion with, or remodelling of, the endosomal mem-
brane. Alternatively, actin removal could be essential for
the effectors to interact with the membrane surface, as
certain protein and phospholipid domains that are cov-
ered in polymerized actin may inhibit their binding to
important regions within the phagosome membrane.
Future perspectives
The study of the molecular basis of Salmonella spp.
pathogenesis in mammals has advanced greatly over
the past 15 years owing to the identification of many
of the key molecules and mechanisms in mammalian
model systems and the discovery of important innate
immune receptors and responses to the bacteria. For
example, important findings have been made in our
understanding of the mechanisms of entrance into non-
phagocytic cells, the way bacteria sense the intra­cellular
environment and remodel their surfaces, and the inflam-
matory responses to salmonellae. Concurrently, the
identification of innate immune receptors for bacterial
products and characterization of ligand binding by these
receptors has progressed greatly. We anticipate a fur-
ther explosion of information in the next decade about
the details of the intracellular lifestyle of salmonellae,
the function of effectors that are translocated across the
phagosome membrane and the response of the host to
the activity of these bacterial proteins. Studying how
salmonellae manipulate endosomal trafficking through
the endocytic pathway should lead to important obser-
vations about unknown mechanisms of endocytic traf-
ficking. In addition, how effectors function to bring
together mammalian protein components that may
not normally be together could offer important find-
ings with relevance for non-infectious diseases. The
 REVIEWS

study of salmonellae in more-resistant mouse models,
coupled with sophisticated imaging, should also lead to
a greater understanding of the pathophysiological role of
identified virulence mechanisms and the development
of chronic disease. This may have relevance for other dis-
eases at mucosal surfaces, such as inflammatory bowel
disease. The future decade may also take the field of
salmonellae pathogenesis and host responses beyond
model systems to the diversity of bacteria and hosts in
nature. Emerging technologies, such as whole-genome
sequencing, will allow us to investigate the diversity
of effectors that are used by the many pathogenic
Salmonella species, contributing to our view of host
specificity. Specific hypotheses will be generated by sta-
tistical association, which will then need to be tested in
model systems. In addition, the availability of human and
animal genotyping will lead to an increase in data about
the diversity of human and animal responses to infec-
tion with various salmonellae. Model systems in various
animals that have diverse innate immune modifications
will also need to be developed to further understand the
natural world in the context of our available tools and
to, ultimately, reach the stage in which the evolution of
microorganisms and the emergence of infectious diseases
can be observed in real time. Therefore, the study of sal-
monellae pathogenesis should yield a rich treasure trove
of information for those who are interested in microbial
pathogenesis, innate immunity, cell biology and genom-
ics, and should remain an important model system of
host–pathogen interactions for many years to come.

1. Pegues, D. A., Ohl, M. E. & Miller, S. I. in Principles
and Practice of Infectious Diseases (eds Mandell,
G. L., Bennet, J. E. & Dolin, R.) 2636–2654 (Churchill
Livingstone, New York, 2005).
2. Garcia-del Portillo, F., Foster, J. W. & Finlay, B. B. Role
of acid tolerance response genes in Salmonella
typhimurium virulence. Infect. Immun. 61, 4489–4492
(1993).
3. Michetti, P., Mahan, M. J., Slauch, J. M., Mekalanos,
J. J. & Neutra, M. R. Monoclonal secretory
immunoglobulin A protects mice against oral
challenge with the invasive pathogen Salmonella
typhimurium. Infect. Immun. 60, 1786–1792 (1992).
4. Selsted, M. E., Miller, S. I., Henschen, A. H. &
Ouellette, A. J. Enteric defensins: antibiotic peptide
components of intestinal host defense. J. Cell Biol.
118, 929–936 (1992).
5. Prouty, A. M., Brodsky, I. E., Falkow, S. & Gunn, J. S.
Bile‑salt‑mediated induction of antimicrobial and bile
resistance in Salmonella typhimurium. Microbiology
150, 775–783 (2004).
6. Francis, C. L., Starnbach, M. N. & Falkow, S.
Morphological and cytoskeletal changes in epithelial
cells occur immediately upon interaction with
Salmonella typhimurium grown under low-oxygen
conditions. Mol. Microbiol. 6, 3077–3087 (1992).
7. Takeuchi, A. Electron microscope studies of
experimental Salmonella infection. I. Penetration into
the intestinal epithelium by Salmonella typhimurium.
Am. J. Pathol. 50, 109–136 (1967).
This reference showed membrane ruffling that is
induced by Salmonella spp. in intestinal cells by
microscopy for the first time.
8. Baumler, A. J., Tsolis, R. M. & Heffron, F. Contribution
of fimbrial operons to attachment to and invasion of
epithelial cell lines by Salmonella typhimurium. Infect.
Immun. 64, 1862–1865 (1996).
9. Kohbata, S., Yokoyama, H. & Yabuuchi, E.
Cytopathogenic effect of Salmonella typhi GIFU 10007
on M cells of murine ileal Peyer’s patches in ligated
ileal loops: an ultrastructural study. Microbiol.
Immunol. 30, 1225–1237 (1986).
10. Jones, B. D., Ghori, N. & Falkow, S. Salmonella
typhimurium initiates murine infection by penetrating
and destroying the specialized epithelial M cells of the
Peyer’s patches. J. Exp. Med. 180, 15–23 (1994).
11. Vazquez-Torres, A. et al. Extraintestinal dissemination
of Salmonella by CD18-expressing phagocytes. Nature
401, 804–808 (1999).
12. Jepson, M. A., Collares-Buzato, C. B., Clark, M. A.,
Hirst, B. H. & Simmons, N. L. Rapid disruption of
epithelial barrier function by Salmonella typhimurium
is associated with structural modification of
intercellular junctions. Infect. Immun. 63, 356–359
(1995).
13. Zhang, S. et al. The Salmonella enterica serotype
Typhimurium effector proteins SipA, SopA, SopB,
SopD, and SopE2 act in concert to induce diarrhea in
calves. Infect. Immun. 70, 3843–3855 (2002).
14. Raffatellu, M. et al. SipA, SopA, SopB, SopD, and
SopE2 contribute to Salmonella enterica serotype
Typhimurium invasion of epithelial cells. Infect. Immun.
73, 146–154 (2005).
15. Stecher, B. et al. Comparison of Salmonella enterica
serovar Typhimurium colitis in germfree mice and mice
pretreated with streptomycin. Infect. Immun. 73,
3228–3241 (2005).
16. Hobbie, S., Chen, L. M., Davis, R. J. & Galan, J. E.
Involvement of mitogen-activated protein kinase
pathways in the nuclear responses and cytokine
production induced by Salmonella typhimurium in
cultured intestinal epithelial cells. J. Immunol. 159,
5550–5559 (1997).
17. Galan, J. E. & Curtiss, R. 3rd Cloning and molecular
characterization of genes whose products allow
Salmonella typhimurium to penetrate tissue culture
cells. Proc. Natl Acad. Sci. USA 86, 6383–6387
(1989).
18. Miller, S. I., Kukral, A. M. & Mekalanos, J. J. A two-
component regulatory system (phoP phoQ) controls
Salmonella typhimurium virulence. Proc. Natl Acad.
Sci. USA 86, 5054–5058 (1989).
This reference identified the PhoP/PhoQ two-
component system as being important for the
regulation of virulence.
19. Alpuche-Aranda, C. M., Swanson, J. A., Loomis, W. P.
& Miller, S. I. Salmonella typhimurium activates
virulence gene transcription within acidified
macrophage phagosomes. Proc. Natl Acad. Sci. USA
89, 10079–10083 (1992).
20. Alpuche-Aranda, C. M., Racoosin, E. L., Swanson, J. A.
& Miller, S. I. Salmonella stimulate macrophage
macropinocytosis and persist within spacious
phagosomes. J. Exp. Med. 179, 601–608 (1994).
21. Ochman, H., Soncini, F. C., Solomon, F. & Groisman,
E. A. Identification of a pathogenicity island required
for Salmonella survival in host cells. Proc. Natl Acad.
Sci. USA 93, 7800–7804 (1996).
References 21 and 22 were the first to show that
the T3SS that is encoded on SPI2 is important for
virulence.
22. Shea, J. E., Hensel, M., Gleeson, C. & Holden, D. W.
Identification of a virulence locus encoding a second
type III secretion system in Salmonella typhimurium.
Proc. Natl Acad. Sci. USA 93, 2593–2597 (1996).
23. Vazquez-Torres, A. et al. Salmonella pathogenicity
island 2‑dependent evasion of the phagocyte NADPH
oxidase. Science 287, 1655–1658 (2000).
24. Miao, E. A., Freeman, J. A. & Miller, S. I. Transcription
of the SsrAB regulon is repressed by alkaline pH and
is independent of PhoPQ and magnesium
concentration. J. Bacteriol. 184, 1493–1497
(2002).
25. Bader, M. W. et al. Regulation of Salmonella
typhimurium virulence gene expression by cationic
antimicrobial peptides. Mol. Microbiol. 50, 219–230
(2003).
26. McCormick, B. A., Miller, S. I., Carnes, D. & Madara,
J. L. Transepithelial signaling to neutrophils by
salmonellae: a novel virulence mechanism for
gastroenteritis. Infect. Immun. 63, 2302–2309
(1995).
27. McCormick, B. A. et al. Surface attachment of
Salmonella typhimurium to intestinal epithelia
imprints the subepithelial matrix with gradients
chemotactic for neutrophils. J. Cell Biol. 131,
1599–1608 (1995).
28. Hansen-Wester, I. & Hensel, M. Salmonella
pathogenicity islands encoding type III secretion
systems. Microbes Infect. 3, 549–559 (2001).
29. Kubori, T. et al. Supramolecular structure of the
Salmonella typhimurium type III protein secretion
system. Science 280, 602–605 (1998).
This study provided the first electron microscopy
images of the T3SS needle complex.
30. Kimbrough, T. G. & Miller, S. I. Contribution of
Salmonella typhimurium type III secretion components
to needle complex formation. Proc. Natl Acad. Sci.
USA 97, 11008–11013 (2000).
31. Kimbrough, T. G. & Miller, S. I. Assembly of the type III
secretion needle complex of Salmonella typhimurium.
Microbes Infect. 4, 75–82 (2002).
32. Miao, E. A. & Miller, S. I. A conserved amino acid
sequence directing intracellular type III secretion by
Salmonella typhimurium. Proc. Natl Acad. Sci. USA
97, 7539–7544 (2000).
33. Brumell, J. H. et al. SopD2 is a novel type III secreted
effector of Salmonella typhimurium that targets late
endocytic compartments upon delivery into host cells.
Traffic 4, 36–48 (2003).
34. Lee, S. H. & Galan, J. E. Salmonella type III secretion-
associated chaperones confer secretion-pathway
specificity. Mol. Microbiol. 51, 483–495 (2004).
35. Karavolos, M. H. et al. Type III secretion of the
Salmonella effector protein SopE is mediated via an
N‑terminal amino acid signal and not an mRNA
sequence. J. Bacteriol. 187, 1559–1567
(2005).
36. Fu, Y. & Galan, J. E. Identification of a specific
chaperone for SptP, a substrate of the centisome 63
type III secretion system of Salmonella typhimurium.
J. Bacteriol. 180, 3393–3399 (1998).
37. Hong, K. H. & Miller, V. L. Identification of a novel
Salmonella invasion locus homologous to Shigella
ipgDE. J. Bacteriol. 180, 1793–1802 (1998).
38. Bronstein, P. A., Miao, E. A. & Miller, S. I. InvB is a
type III secretion chaperone specific for SspA.
J. Bacteriol. 182, 6638–6644 (2000).
39. Tucker, S. C. & Galan, J. E. Complex function for SicA,
a Salmonella enterica serovar Typhimurium type III
secretion-associated chaperone. J. Bacteriol. 182,
2262–2268 (2000).
40. Ehrbar, K., Friebel, A., Miller, S. I. & Hardt, W. D.
Role of the Salmonella pathogenicity island 1
(SPI‑1) protein InvB in type III secretion of SopE and
SopE2, two Salmonella effector proteins encoded
outside of SPI‑1. J. Bacteriol. 185, 6950–6967
(2003).
41. Higashide, W. & Zhou, D. The first 45 amino acids of
SopA are necessary for InvB binding and SPI‑1
secretion. J. Bacteriol. 188, 2411–2420 (2006).
42. Akeda, Y. & Galan, J. E. Chaperone release and
unfolding of substrates in type III secretion. Nature
437, 911–915 (2005).
43. Brown, N. F. et al. Salmonella pathogenicity island 2 is
expressed prior to penetrating the intestine. PLoS
Pathog. 1, e32 (2005).
44. Hensel, M. et al. Functional analysis of ssaJ and the
ssaK/U operon, 13 genes encoding components of the
type III secretion apparatus of Salmonella pathogenicity
island 2. Mol. Microbiol. 24, 155–167 (1997).
45. Deiwick, J. et al. Mutations in Salmonella pathogenicity
island 2 (SPI2) genes affecting transcription of SPI1
genes and resistance to antimicrobial agents.
J. Bacteriol. 180, 4775–4780 (1998).

nature reviews | microbiology volume 6 | january 2008 | 63

© 2008 Nature Publishing Group
REVIEWS
46. Steele-Mortimer, O. et al. The invasion-associated
type III secretion system of Salmonella enterica
serovar Typhimurium is necessary for intracellular
proliferation and vacuole biogenesis in epithelial cells.
Cell. Microbiol. 4, 43–54 (2002).
47. Hernandez, L. D., Hueffer, K., Wenk, M. R. & Galan,
J. E. Salmonella modulates vesicular traffic by
altering phosphoinositide metabolism. Science 304,
1805–1807 (2004).
48. Drecktrah, D., Knodler, L. A., Galbraith, K. & Steele-
Mortimer, O. The Salmonella SPI1 effector SopB
stimulates nitric oxide production long after invasion.
Cell. Microbiol. 7, 105–113 (2005).
49. Lawley, T. D. et al. Genome-wide screen for Salmonella
genes required for long-term systemic infection of the
mouse. PLoS Pathog. 2, e11 (2006).
50. Brawn, L. C., Hayward, R. D. & Koronakis, V.
Salmonella SPI1 effector SipA persists after entry and
cooperates with a SPI2 effector to regulate
phagosome maturation and intracellular replication.
Cell Host Microbe 1, 63–75 (2007).
51. Giacomodonato, M. N. et al. SipA, SopA, SopB, SopD
and SopE2 effector proteins of Salmonella enterica
serovar Typhimurium are synthesized at late stages of
infection in mice. Microbiology 153, 1221–1228
(2007).
52. Li, J. et al. Relationship between evolutionary rate and
cellular location among the Inv/Spa invasion proteins
of Salmonella enterica. Proc. Natl Acad. Sci. USA 92,
7252–7256 (1995).
53. Ochman, H. & Groisman, E. A. Distribution of
pathogenicity islands in Salmonella spp. Infect.
Immun. 64, 5410–5412 (1996).
54. Boyd, E. F., Li, J., Ochman, H. & Selander, R. K.
Comparative genetics of the inv-spa invasion gene
complex of Salmonella enterica. J. Bacteriol. 179,
1985–1991 (1997).
55. Hensel, M. et al. Analysis of the boundaries of
Salmonella pathogenicity island 2 and the
corresponding chromosomal region of Escherichia coli
K‑12. J. Bacteriol. 179, 1105–1111 (1997).
56. Hensel, M., Nikolaus, T. & Egelseer, C. Molecular and
functional analysis indicates a mosaic structure of
Salmonella pathogenicity island 2. Mol. Microbiol. 31,
489–498 (1999).
57. Porwollik, S., Wong, R. M. & McClelland, M.
Evolutionary genomics of Salmonella: gene
acquisitions revealed by microarray analysis.
Proc. Natl Acad. Sci. USA 99, 8956–8961 (2002).
58. Hormaeche, C. E. Natural resistance to Salmonella
typhimurium in different inbred mouse strains.
Immunology 37, 311–318 (1979).
59. Vidal, S. M., Malo, D., Vogan, K., Skamene, E. &
Gros, P. Natural resistance to infection with
intracellular parasites: isolation of a candidate for Bcg.
Cell 73, 469–485 (1993).
60. Hensel, M. et al. Genes encoding putative effector
proteins of the type III secretion system of
Salmonella pathogenicity island 2 are required for
bacterial virulence and proliferation in macrophages.
Mol. Microbiol. 30, 163–174 (1998).
61. Beuzon, C. R. & Holden, D. W. Use of mixed
infections with Salmonella strains to study virulence
genes and their interactions in vivo. Microbes Infect.
3, 1345–1352 (2001).
62. Monack, D. M., Bouley, D. M. & Falkow, S. Salmonella
typhimurium persists within macrophages in the
mesenteric lymph nodes of chronically infected
Nramp1+/+ mice and can be reactivated by IFNgamma
neutralization. J. Exp. Med. 199, 231–241 (2004).
63. Behlau, I. & Miller, S. I. A PhoP-repressed gene
promotes Salmonella typhimurium invasion of
epithelial cells. J. Bacteriol. 175, 4475–4484 (1993).
64. Barthel, M. et al. Pretreatment of mice with
streptomycin provides a Salmonella enterica serovar
Typhimurium colitis model that allows analysis of both
pathogen and host. Infect. Immun. 71, 2839–2858
(2003).
65. Hardt, W. D., Chen, L. M., Schuebel, K. E., Bustelo,
X. R. & Galan, J. E. S. typhimurium encodes an
activator of Rho GTPases that induces membrane
ruffling and nuclear responses in host cells. Cell 93,
815–826 (1998).
This work elegantly demonstrated the ability of
SopE to activate GTPases and subsequent actin
rearrangements and transcriptional responses.
66. Bakshi, C. S. et al. Identification of SopE2, a
Salmonella secreted protein which is highly
homologous to SopE and involved in bacterial invasion
of epithelial cells. J. Bacteriol. 182, 2341–2344
(2000).
64 | january 2008 | volume 6 www.nature.com/reviews/micro
67. Stender, S. et al. Identification of SopE2 from
Salmonella typhimurium, a conserved guanine
nucleotide exchange factor for Cdc42 of the host cell.
Mol. Microbiol. 36, 1206–1221 (2000).
68. Friebel, A. et al. SopE and SopE2 from Salmonella
typhimurium activate different sets of RhoGTPases of
the host cell. J. Biol. Chem. 276, 34035–34040
(2001).
69. Zhou, D., Chen, L. M., Hernandez, L., Shears, S. B. &
Galan, J. E. A Salmonella inositol polyphosphatase
acts in conjunction with other bacterial effectors to
promote host cell actin cytoskeleton rearrangements
and bacterial internalization. Mol. Microbiol. 39,
248–259 (2001).
70. Patel, J. C. & Galan, J. E. Differential activation and
function of Rho GTPases during Salmonella-host cell
interactions. J. Cell Biol. 175, 453–463 (2006).
71. Ellerbroek, S. M. et al. SGEF, a RhoG guanine
nucleotide exchange factor that stimulates
macropinocytosis. Mol. Biol. Cell 15, 3309–3319
(2004).
72. Criss, A. K. & Casanova, J. E. Coordinate regulation of
Salmonella enterica serovar Typhimurium invasion of
epithelial cells by the Arp2/3 complex and Rho
GTPases. Infect. Immun. 71, 2885–2891 (2003).
73. Unsworth, K. E., Way, M., McNiven, M., Machesky, L.
& Holden, D. W. Analysis of the mechanisms of
Salmonella-induced actin assembly during invasion of
host cells and intracellular replication. Cell. Microbiol.
6, 1041–1055 (2004).
74. Shi, J., Scita, G. & Casanova, J. E. WAVE2 signaling
mediates invasion of polarized epithelial cells by
Salmonella typhimurium. J. Biol. Chem. 280,
29849–29855 (2005).
75. Hayward, R. D. & Koronakis, V. Direct nucleation and
bundling of actin by the SipC protein of invasive
Salmonella. EMBO J. 18, 4926–4934 (1999).
References 75 and 76 detected direct
manipulation of actin by the SPI1 T3SS effectors
SipA and SipC.
76. Zhou, D., Mooseker, M. S. & Galan, J. E. Role of the
S. typhimurium actin-binding protein SipA in bacterial
internalization. Science 283, 2092–2095 (1999).
77. Scherer, C. A., Cooper, E. & Miller, S. I. The Salmonella
type III secretion translocon protein SspC is inserted
into the epithelial cell plasma membrane upon
infection. Mol. Microbiol. 37, 1133–1145 (2000).
78. McGhie, E. J., Hayward, R. D. & Koronakis, V.
Cooperation between actin-binding proteins of
invasive Salmonella: SipA potentiates SipC nucleation
and bundling of actin. EMBO J. 20, 2131–2139
(2001).
79. Higashide, W., Dai, S., Hombs, V. P. & Zhou, D.
Involvement of SipA in modulating actin dynamics
during Salmonella invasion into cultured epithelial
cells. Cell. Microbiol. 4, 357–365 (2002).
80. Chen, L. M., Hobbie, S. & Galan, J. E. Requirement
of CDC42 for Salmonella-induced cytoskeletal and
nuclear responses. Science 274, 2115–2118
(1996).
81. Boyle, E. C., Brown, N. F. & Finlay, B. B. Salmonella
enterica serovar Typhimurium effectors SopB, SopE,
SopE2 and SipA disrupt tight junction structure and
function. Cell. Microbiol. 8, 1946–1957 (2006).
82. Norris, F. A., Wilson, M. P., Wallis, T. S., Galyov, E. E. &
Majerus, P. W. SopB, a protein required for virulence
of Salmonella dublin, is an inositol phosphate
phosphatase. Proc. Natl Acad. Sci. USA 95,
14057–14059 (1998).
83. Hersh, D. et al. The Salmonella invasin SipB induces
macrophage apoptosis by binding to caspase‑1.
Proc. Natl. Acad. Sci. USA 96, 2396–2401 (1999).
84. Franchi, L. et al. Cytosolic flagellin requires Ipaf for
activation of caspase‑1 and interleukin 1b in
Salmonella-infected macrophages. Nature Immunol. 7,
576–582 (2006).
85. Miao, E. A. et al. Cytoplasmic flagellin activates
caspase‑1 and secretion of interleukin 1b via Ipaf.
Nature Immunol. 7, 569–575 (2006).
86. Lara-Tejero, M. et al. Role of the caspase‑1
inflammasome in Salmonella typhimurium
pathogenesis. J. Exp. Med. 203, 1407–1412 (2006).
87. Raupach, B., Peuschel, S. K., Monack, D. M. &
Zychlinsky, A. Caspase‑1‑mediated activation of
interleukin-1b (IL-1b) and IL‑18 contributes to innate
immune defenses against Salmonella enterica serovar
Typhimurium infection. Infect. Immun. 74, 4922–4926
(2006).
88. Jones, M. A. et al. Secreted effector proteins of
Salmonella dublin act in concert to induce enteritis.
Infect. Immun. 66, 5799–5804 (1998).
© 2008 Nature Publishing Group
89. Wood, M. W. et al. The secreted effector protein of
Salmonella dublin, SopA, is translocated into
eukaryotic cells and influences the induction of
enteritis. Cell Microbiol. 2, 293–303 (2000).
90. Zhang, Y., Higashide, W. M., McCormick, B. A., Chen, J.
& Zhou, D. The inflammation-associated Salmonella
SopA is a HECT-like E3 ubiquitin ligase. Mol. Microbiol.
62, 786–793 (2006).
91. Jiang, X. et al. The related effector proteins SopD and
SopD2 from Salmonella enterica serovar Typhimurium
contribute to virulence during systemic infection of
mice. Mol. Microbiol. 54, 1186–1198 (2004).
92. Fu, Y. & Galan, J. E. A Salmonella protein
antagonizes Rac‑1 and Cdc42 to mediate host-cell
recovery after bacterial invasion. Nature 401,
293–297 (1999).
References 92 and 93 identified the
GTPase-antagonizing activity of SptP and showed
how the activities of SopE and SptP are temporally
regulated by proteasome-dependent degradation.
93. Kubori, T. & Galan, J. E. Temporal regulation of
Salmonella virulence effector function by proteasome-
dependent protein degradation. Cell 115, 333–342
(2003).
94. Murli, S., Watson, R. O. & Galan, J. E. Role of tyrosine
kinases and the tyrosine phosphatase SptP in the
interaction of Salmonella with host cells. Cell.
Microbiol. 3, 795–810 (2001).
95. Haraga, A. & Miller, S. I. A Salmonella enterica
serovar Typhimurium translocated leucine-rich repeat
effector protein inhibits NF‑kappa B‑dependent gene
expression. Infect. Immun. 71, 4052–4058 (2003).
96. Miao, E. A. et al. Salmonella typhimurium leucine-
rich repeat proteins are targeted to the SPI1 and
SPI2 type III secretion systems. Mol. Microbiol. 34,
850–864 (1999).
97. Rohde, J. R., Breitkreutz, A., Chenal, A., Sansonetti,
P. J. & Parsot, C. Type III secretion effectors of the
IpaH family are E3 ubiquitin ligases. Cell Host Microbe
1, 77–83 (2007).
98. Haraga, A. & Miller, S. I. A Salmonella type III
secretion effector interacts with the mammalian
serine/threonine protein kinase PKN1. Cell. Microbiol.
8, 837–846 (2006).
99. Collier-Hyams, L. S. et al. Cutting edge: Salmonella
AvrA effector inhibits the key proinflammatory, anti-
apoptotic NF‑kappaB pathway. J. Immunol. 169,
2846–2850 (2002).
100. Mittal, R., Peak-Chew, S. Y. & McMahon, H. T.
Acetylation of MEK2 and I kappa B kinase (IKK)
activation loop residues by YopJ inhibits signaling.
Proc. Natl Acad. Sci. USA 103, 18574–18579
(2006).
101. Mukherjee, S. et al. Yersinia YopJ acetylates and
inhibits kinase activation by blocking phosphorylation.
Science 312, 1211–1214 (2006).
102. Buchwald, D. S. & Blaser, M. J. A review of human
salmonellosis: II. Duration of excretion following
infection with nontyphi Salmonella. Rev. Infect. Dis. 6,
345–356 (1984).
103. Richter-Dahlfors, A., Buchan, A. M. & Finlay, B. B.
Murine salmonellosis studied by confocal microscopy:
Salmonella typhimurium resides intracellularly inside
macrophages and exerts a cytotoxic effect on
phagocytes in vivo. J. Exp. Med. 186, 569–580
(1997).
104. Fields, P. I., Swanson, R. V., Haidaris, C. G. & Heffron, F.
Mutants of Salmonella typhimurium that cannot survive
within the macrophage are avirulent. Proc. Natl Acad.
Sci. USA 83, 5189–5193 (1986).
This important study demonstrated that survival
within macrophages is important for S. typhimurium
virulence.
105. O’Callaghan, D., Maskell, D., Liew, F. Y., Easmon, C. S.
& Dougan, G. Characterization of aromatic- and
purine-dependent Salmonella typhimurium: attention,
persistence, and ability to induce protective immunity
in BALB/c mice. Infect. Immun. 56, 419–423 (1988).
106. Dukes, J. D. et al. The secreted Salmonella dublin
phosphoinositide phosphatase, SopB, localizes to
PtdIns(3)P-containing endosomes and perturbs
normal endosome to lysosome trafficking. Biochem. J.
395, 239–247 (2006).
107. Carrol, M. E., Jackett, P. S., Aber, V. R. & Lowrie,
D. B. Phagolysosome formation, cyclic adenosine
3′:5′-monophosphate and the fate of Salmonella
typhimurium within mouse peritoneal macrophages.
J. Gen. Microbiol. 110, 421–429 (1979).
108. Buchmeier, N. A. & Heffron, F. Inhibition of macrophage
phagosome–lysosome fusion by Salmonella
typhimurium. Infect. Immun. 59, 2232–2238 (1991).

109. Oh, Y. K. et al. Rapid and complete fusion of
macrophage lysosomes with phagosomes containing
Salmonella typhimurium. Infect. Immun. 64,
3877–3883 (1996).
110. Drecktrah, D., Knodler, L. A., Howe, D. & Steele-
Mortimer, O. Salmonella trafficking is defined by
continuous dynamic interactions with the
endolysosomal system. Traffic 8, 212–225 (2007).
111. Rathman, M., Sjaastad, M. D. & Falkow, S.
Acidification of phagosomes containing Salmonella
typhimurium in murine macrophages. Infect. Immun.
64, 2765–2773 (1996).
112. Martin-Orozco, N. et al. Visualization of vacuolar
acidification-induced transcription of genes of
pathogens inside macrophages. Mol. Biol. Cell 17,
498–510 (2006).
113. Steele-Mortimer, O., Meresse, S., Gorvel, J. P., Toh,
B. H. & Finlay, B. B. Biogenesis of Salmonella
typhimurium-containing vacuoles in epithelial cells
involves interactions with the early endocytic pathway.
Cell. Microbiol. 1, 33–49 (1999).
114. Smith, A. C., Cirulis, J. T., Casanova, J. E., Scidmore,
M. A. & Brumell, J. H. Interaction of the Salmonella-
containing vacuole with the endocytic recycling
system. J. Biol. Chem. 280, 24634–24641 (2005).
115. Garcia-del Portillo, F. & Finlay, B. B. Targeting of
Salmonella typhimurium to vesicles containing
lysosomal membrane glycoproteins bypasses
compartments with mannose 6‑phosphate receptors.
J. Cell Biol. 129, 81–97 (1995).
116. Meresse, S., Steele-Mortimer, O., Finlay, B. B. &
Gorvel, J. P. The rab7 GTPase controls the maturation
of Salmonella typhimurium-containing vacuoles in
HeLa cells. EMBO J. 18, 4394–4403 (1999).
117. Hashim, S., Mukherjee, K., Raje, M., Basu, S. K. &
Mukhopadhyay, A. Live Salmonella modulate
expression of Rab proteins to persist in a specialized
compartment and escape transport to lysosomes.
J. Biol. Chem. 275, 16281–16288 (2000).
118. Brumell, J. H., Tang, P., Mills, S. D. & Finlay, B. B.
Characterization of Salmonella-induced filaments (Sifs)
reveals a delayed interaction between Salmonella-
containing vacuoles and late endocytic compartments.
Traffic 2, 643–653 (2001).
119. Garvis, S. G., Beuzon, C. R. & Holden, D. W. A role for
the PhoP/Q regulon in inhibition of fusion between
lysosomes and Salmonella-containing vacuoles in
macrophages. Cell. Microbiol. 3, 731–744 (2001).
120. Cuellar-Mata, P. et al. Nramp1 modifies the fusion of
Salmonella typhimurium-containing vacuoles with
cellular endomembranes in macrophages. J. Biol.
Chem. 277, 2258–2265 (2002).
121. Catron, D. M. et al. The Salmonella-containing vacuole
is a major site of intracellular cholesterol accumulation
and recruits the GPI-anchored protein CD55. Cell.
Microbiol. 4, 315–328 (2002).
122. Fields, P. I., Groisman, E. A. & Heffron, F. A Salmonella
locus that controls resistance to microbicidal proteins
from phagocytic cells. Science 243, 1059–1062
(1989).
123. Miller, S. I. & Mekalanos, J. J. Constitutive expression
of the PhoP regulon attenuates Salmonella virulence
and survival within macrophages. J. Bacteriol. 172,
2485–2490 (1990).
124. Nickerson, C. A. & Curtiss, R. Role of sigma factor
RpoS in initial stages of Salmonella typhimurium
infection. Infect. Immun. 65, 1814–1823 (1997).
125. Bearson, B. L., Wilson, L. & Foster, J. W. A low pH-
inducible, PhoPQ-dependent acid tolerance response
protects Salmonella typhimurium against inorganic
acid stress. J. Bacteriol. 180, 2409–2417 (1998).
126. Shiloh, M. U. et al. Phenotype of mice and
macrophages deficient in both phagocyte oxidase and
inducible nitric oxide synthase. Immunity 10, 29–38
(1999).
127. Gunn, J. S., Ryan, S. S., Van Velkinburgh, J. C.,
Ernst, R. K. & Miller, S. I. Genetic and functional
analysis of a PmrA–PmrB‑regulated locus necessary
for lipopolysaccharide modification, antimicrobial
peptide resistance, and oral virulence of Salmonella
enterica serovar Typhimurium. Infect. Immun. 68,
6139–6146 (2000).
128. Vazquez-Torres, A., Jones-Carson, J., Mastroeni, P.,
Ischiropoulos, H. & Fang, F. C. Antimicrobial actions of
the NADPH phagocyte oxidase and inducible nitric
oxide synthase in experimental salmonellosis. I.
Effects on microbial killing by activated peritoneal
macrophages in vitro. J. Exp. Med. 192, 227–236
(2000).
129. Hisert, K. B. et al. A glutamate‑alanine‑leucine (EAL)
domain protein of Salmonella controls bacterial
nature reviews | microbiology volume 6 | january 2008 | 65
survival in mice, antioxidant defence and killing of
macrophages: role of cyclic diGMP. Mol. Microbiol.
56, 1234–1245 (2005).
130. Humphreys, S., Stevenson, A., Bacon, A., Weinhardt,
A. B. & Roberts, M. The alternative sigma factor,
sigmaE, is critically important for the virulence of
Salmonella typhimurium. Infect. Immun. 67,
1560–1568 (1999).
131. Rosenberger, C. M., Gallo, R. L. & Finlay, B. B.
Interplay between antibacterial effectors: a
macrophage antimicrobial peptide impairs
intracellular Salmonella replication. Proc. Natl Acad.
Sci. USA 101, 2422–2427 (2004).
132. Curtiss, R. & Kelly, S. M. Salmonella typhimurium
deletion mutants lacking adenylate cyclase and cyclic
AMP receptor protein are avirulent and immunogenic.
Infect. Immun. 55, 3035–3043 (1987).
133. Fang, F. C. et al. The alternative sigma factor katF
(rpoS) regulates Salmonella virulence. Proc. Natl Acad.
Sci. USA 89, 11978–11982 (1992).
134. Lee, A. K., Detweiler, C. S. & Falkow, S. OmpR
regulates the two-component system SsrA-SsrB in
Salmonella pathogenicity island 2. J. Bacteriol. 182,
771–781 (2000).
135. Testerman, T. L. et al. The alternative sigma factor sE
controls antioxidant defences required for Salmonella
virulence and stationary-phase survival. Mol.
Microbiol. 43, 771–782 (2002).
136. Prost, L. R. et al. Activation of the bacterial sensor
kinase PhoQ by acidic pH. Mol. Cell 26, 165–174
(2007).
References 136 and 138 show that PhoQ directly
senses antimicrobial peptides and low pH.
137. Heithoff, D. M. et al. Coordinate intracellular
expression of Salmonella genes induced during
infection. J. Bacteriol. 181, 799–807 (1999).
138. Bader, M. W. et al. Recognition of antimicrobial
peptides by a bacterial sensor kinase. Cell 122,
461–472 (2005).
139. Cho, U. S. et al. Metal bridges between the PhoQ
sensor domain and the membrane regulate
transmembrane signaling. J. Mol. Biol. 356,
1193–1206 (2006).
140. Gibbons, H. S., Kalb, S. R., Cotter, R. J. & Raetz, C. R.
Role of Mg2+ and pH in the modification of Salmonella
lipid A after endocytosis by macrophage tumour cells.
Mol. Microbiol. 55, 425–440 (2005).
141. Ernst, R. K., Guina, T. & Miller, S. I. Salmonella
typhimurium outer membrane remodeling: role in
resistance to host innate immunity. Microbes Infect. 3,
1327–1334 (2001).
142. Guo, L. et al. Regulation of lipid A modifications by
Salmonella typhimurium virulence genes phoP-phoQ.
Science 276, 250–253 (1997).
This study demonstrates that PhoP/PhoQ regulates
the remodelling of the LPS structure in a way that
makes it less immunostimulatory by TLR4 signalling.
143. Gunn, J. S. et al. PmrA‑PmrB‑regulated genes
necessary for 4‑aminoarabinose lipid A modification
and polymyxin resistance. Mol. Microbiol. 27,
1171–1182 (1998).
144. Guo, L. et al. Lipid A acylation and bacterial resistance
against vertebrate antimicrobial peptides. Cell 95,
189–198 (1998).
145. Hilbert, F., Garcia-del Portillo, F. & Groisman, E. A.
A periplasmic d‑alanyl‑d-alanine dipeptidase in the
Gram-negative bacterium Salmonella enterica.
J. Bacteriol. 181, 2158–2165 (1999).
146. Fang, F. C. et al. Virulent Salmonella typhimurium has
two periplasmic Cu, Zn-superoxide dismutases. Proc.
Natl Acad. Sci. USA 96, 7502–7507 (1999).
147. Eriksson, S., Lucchini, S., Thompson, A., Rhen, M. &
Hinton, J. C. Unravelling the biology of macrophage
infection by gene expression profiling of intracellular
Salmonella enterica. Mol. Microbiol. 47, 103–118
(2003).
148. Cirillo, D. M., Valdivia, R. H., Monack, D. M. & Falkow,
S. Macrophage-dependent induction of the Salmonella
pathogenicity island 2 type III secretion system and its
role in intracellular survival. Mol. Microbiol. 30,
175–188 (1998).
149. Hensel, M. et al. Simultaneous identification of
bacterial virulence genes by negative selection.
Science 269, 400–403 (1995).
150. Uchiya, K. et al. A Salmonella virulence protein that
inhibits cellular trafficking. EMBO J. 18, 3924–3933
(1999).
151. Shotland, Y., Kramer, H. & Groisman, E. A. The
Salmonella SpiC protein targets the mammalian
Hook3 protein function to alter cellular trafficking.
Mol. Microbiol. 49, 1565–1576 (2003).
© 2008 Nature Publishing Group
REVIEWS
152. Freeman, J. A., Rappl, C., Kuhle, V., Hensel, M. &
Miller, S. I. SpiC is required for translocation of
Salmonella pathogenicity island 2 effectors and
secretion of translocon proteins SseB and SseC.
J. Bacteriol. 184, 4971–4980 (2002).
153. Yu, X. J. et al. SpiC is required for secretion of
Salmonella pathogenicity island 2 type III secretion
system proteins. Cell. Microbiol. 4, 531–540 (2002).
154. Beuzon, C. R. et al. Salmonella maintains the
integrity of its intracellular vacuole through the
action of SifA. EMBO J. 19, 3235–3249 (2000).
155. Ruiz-Albert, J. et al. Complementary activities of SseJ
and SifA regulate dynamics of the Salmonella
typhimurium vacuolar membrane. Mol. Microbiol. 44,
645–661 (2002).
156. Freeman, J. A., Ohl, M. E. & Miller, S. I. The
Salmonella enterica serovar Typhimurium translocated
effectors SseJ and SifsB are targeted to the
Salmonella-containing vacuole. Infect. Immun. 71,
418–427 (2003).
157. Knodler, L. A. et al. Salmonella type III effectors PipB
and PipB2 are targeted to detergent-resistant
microdomains on internal host cell membranes. Mol.
Microbiol. 49, 685–704 (2003).
158. Deiwick, J. et al. The translocated Salmonella effector
proteins SseF and SseG interact and are required to
establish an intracellular replication niche. Infect.
Immun. 74, 6965–6972 (2006).
159. Brumell, J. H., Goosney, D. L. & Finlay, B. B. SifA, a
type III secreted effector of Salmonella typhimurium,
directs Salmonella-induced filament (Sif) formation
along microtubules. Traffic 3, 407–415 (2002).
160. Miao, E. A. et al. Salmonella effectors translocated
across the vacuolar membrane interact with the
actin cytoskeleton. Mol. Microbiol. 48, 401–415
(2003).
161. Salcedo, S. P. & Holden, D. W. SseG, a virulence
protein that targets Salmonella to the Golgi network.
EMBO J. 22, 5003–5014 (2003).
162. Kuhle, V., Jackel, D. & Hensel, M. Effector proteins
encoded by Salmonella pathogenicity island 2
interfere with the microtubule cytoskeleton after
translocation into host cells. Traffic 5, 356–370
(2004).
163. Garcia-del Portillo, F., Zwick, M. B., Leung, K. Y. &
Finlay, B. B. Salmonella induces the formation of
filamentous structures containing lysosomal
membrane glycoproteins in epithelial cells.
Proc. Natl Acad. Sci. USA 90, 10544–10548 (1993).
References 163 and 164 were the first to show
that S. typhimurium produces Sifs in infected
cultured cells and that the effector that is
responsible for this activity is SifA.
164. Stein, M. A., Leung, K. Y., Zwick, M., Garcia-del
Portillo, F. & Finlay, B. B. Identification of a Salmonella
virulence gene required for formation of filamentous
structures containing lysosomal membrane
glycoproteins within epithelial cells. Mol. Microbiol.
20, 151–164 (1996).
165. Knodler, L. A. & Steele-Mortimer, O. The Salmonella
effector PipB2 affects late endosome/lysosome
distribution to mediate Sif extension. Mol. Biol. Cell.
16, 4108–4123 (2005).
166. Boucrot, E., Henry, T., Borg, J. P., Gorvel, J. P. &
Meresse, S. The intracellular fate of Salmonella
depends on the recruitment of kinesin. Science 308,
1174–1178 (2005).
167. Alto, N. M. et al. Identification of a bacterial type III
effector family with G protein mimicry functions. Cell
124, 133–145 (2006).
168. Reinicke, A. T. et al. A Salmonella typhimurium
effector protein SifA is modified by host cell
prenylation and S‑acylation machinery. J. Biol. Chem.
280, 14620–14627 (2005).
169. Guy, R. L., Gonias, L. A. & Stein, M. A. Aggregation of
host endosomes by Salmonella requires SPI2
translocation of SseFG and involves SpvR and the fms-
aroE intragenic region. Mol. Microbiol. 37,
1417–1435 (2000).
170. Henry, T. et al. The Salmonella effector protein PipB2
is a linker for kinesin‑1. Proc. Natl Acad. Sci. USA
103, 13497–13502 (2006).
171. Hansen-Wester, I., Stecher, B. & Hensel, M. Type III
secretion of Salmonella enterica serovar Typhimurium
translocated effectors and SseFG. Infect. Immun. 70,
1403–1409 (2002).
172. Kuhle, V. & Hensel, M. SseF and SseG are
translocated effectors of the type III secretion system
of Salmonella pathogenicity island 2 that modulate
aggregation of endosomal compartments. Cell.
Microbiol. 4, 813–824 (2002).
REVIEWS
173. Abrahams, G. L., Muller, P. & Hensel, M. Functional
dissection of SseF, a type III effector protein involved
in positioning the Salmonella-containing vacuole.
Traffic 7, 950–965 (2006).
174. Kuhle, V., Abrahams, G. L. & Hensel, M. Intracellular
Salmonella enterica redirect exocytic transport
processes in a Salmonella pathogenicity island
2‑dependent manner. Traffic 7, 716–730 (2006).
175. Birmingham, C. L., Jiang, X., Ohlson, M. B., Miller, S. I.
& Brumell, J. H. Salmonella-induced filament formation
is a dynamic phenotype induced by rapidly replicating
Salmonella enterica serovar Typhimurium in epithelial
cells. Infect. Immun. 73, 1204–1208 (2005).
176. Lesnick, M. L., Reiner, N. E., Fierer, J. & Guiney, D. G.
The Salmonella spvB virulence gene encodes an
enzyme that ADP-ribosylates actin and destabilizes
the cytoskeleton of eukaryotic cells. Mol. Microbiol.
39, 1464–1470 (2001).
177. Browne, S. H., Lesnick, M. L. & Guiney, D. G. Genetic
requirements for Salmonella-induced cytopathology in
human monocyte-derived macrophages. Infect.
Immun. 70, 7126–7135 (2002).
178. Gotoh, H. et al. Extracellular secretion of the virulence
plasmid-encoded ADP-ribosyltransferase SpvB in
Salmonella. Microb. Pathog. 34, 227–238 (2003).
179. Brumlik, M. J. & Buckley, J. T. Identification of the
catalytic triad of the lipase/acyltransferase from
Aeromonas hydrophila. J. Bacteriol. 178, 2060–2064
(1996).
180. Ohlson, M. B., Fluhr, K., Birmingham, C. L., Brumell,
J. H. & Miller, S. I. SseJ deacylase activity by
Salmonella enterica serovar Typhimurium promotes
virulence in mice. Infect. Immun. 73, 6249–6259
(2005).
181. Harrison, R. E. et al. Salmonella impairs RILP
recruitment to Rab7 during maturation of invasion
vacuoles. Mol. Biol. Cell 15, 3146–3154 (2004).
182. Marsman, M., Jordens, I., Kuijl, C., Janssen, L. &
Neefjes, J. Dynein-mediated vesicle transport controls
intracellular Salmonella replication. Mol. Biol. Cell 15,
2954–2964 (2004).
183. Meresse, S. et al. Remodelling of the actin cytoskeleton
is essential for replication of intravacuolar Salmonella.
Cell. Microbiol. 3, 567–577 (2001).
184. Viboud, G. I. & Bliska, J. B. A bacterial type III
secretion system inhibits actin polymerization to
prevent pore formation in host cell membranes.
EMBO J. 20, 5373–5382 (2001).
66 | january 2008 | volume 6 www.nature.com/reviews/micro
185. Marlovits, T. C. et al. Structural insights into the
assembly of the type III secretion needle complex.
Science 306, 1040–1042 (2004).
186. Sun, J., Hobert, M. E., Rao, A. S., Neish, A. S. &
Madara, J. L. Bacterial activation of b-catenin
signaling in human epithelia. Am. J. Physiol.
Gastrointest. Liver Physiol. 287, G220–227 (2004).
187. Zhou, D., Mooseker, M. S. & Galan, J. E. An invasion-
associated Salmonella protein modulates the actin-
bundling activity of plastin. Proc. Natl Acad. Sci. USA
96, 10176–10181 (1999).
188. Lee, C. A. et al. A secreted Salmonella protein induces
a proinflammatory response in epithelial cells, which
promotes neutrophil migration. Proc. Natl Acad. Sci.
USA 97, 12283–12288 (2000).
189. Hernandez, L. D., Pypaert, M., Flavell, R. A. & Galan,
J. E. A Salmonella protein causes macrophage cell
death by inducing autophagy. J. Cell Biol. 163,
1123–1131 (2003).
190. Hayward, R. D. et al. Cholesterol binding by the
bacterial type III translocon is essential for virulence
effector delivery into mammalian cells. Mol. Microbiol.
56, 590–603 (2005).
191. Carlson, S. A., Omary, M. B. & Jones, B. D.
Identification of cytokeratins as accessory mediators of
Salmonella entry into eukaryotic cells. Life Sci. 70,
1415–1426 (2002).
192. Knodler, L. A., Finlay, B. B. & Steele-Mortimer, O. The
Salmonella effector protein SopB protects epithelial
cells from apoptosis by sustained activation of Akt.
J. Biol. Chem. 280, 9058–9064 (2005).
193. Mukherjee, K., Parashuraman, S., Raje, M. &
Mukhopadhyay, A. SopE acts as an Rab5-specific
nucleotide exchange factor and recruits non-
prenylated Rab5 on Salmonella-containing
phagosomes to promote fusion with early endosomes.
J. Biol. Chem. 276, 23607–23615 (2001).
194. Coombes, B. K. et al. Genetic and molecular analysis
of GogB, a phage-encoded type III-secreted substrate
in Salmonella enterica serovar Typhimurium with
autonomous expression from its associated phage.
J. Mol. Biol. 348, 817–830 (2005).
195. Brumell, J. H., Rosenberger, C. M., Gotto, G. T.,
Marcus, S. L. & Finlay, B. B. SifA permits survival
and replication of Salmonella typhimurium in murine
macrophages. Cell. Microbiol. 3, 75–84 (2001).
196. Tezcan-Merdol, D. et al. Actin is ADP-ribosylated by
the Salmonella enterica virulence-associated protein
© 2008 Nature Publishing Group
SpvB. Mol. Microbiol. 39, 606–619 (2001).
197. Worley, M. J., Nieman, G. S., Geddes, K. & Heffron, F.
Salmonella typhimurium disseminates within its host
by manipulating the motility of infected cells. Proc.
Natl Acad. Sci. USA 103, 17915–17920 (2006).
198. Kujat Choy, S. L. et al. SseK1 and SseK2 are novel
translocated proteins of Salmonella enterica serovar
Typhimurium. Infect. Immun. 72, 5115–5125
(2004).
199. Rytkonen, A. et al. SseL, a Salmonella deubiquitinase
required for macrophage killing and virulence. Proc.
Natl Acad. Sci. USA 104, 3502–3507 (2007).
200. Geddes, K., Worley, M., Niemann, G. & Heffron, F.
Identification of new secreted effectors in Salmonella
enterica serovar Typhimurium. Infect. Immun. 73,
6260–6271 (2005).
201. Tsolis, R. M., Adams, L. G., Ficht, T. A. & Baumler, A. J.
Contribution of Salmonella typhimurium virulence
factors to diarrheal disease in calves. Infect. Immun.
67, 4879–4885 (1999).
Acknowledgments
We would like to thank the entire Salmonella pathogenesis
community for its work that made this Review possible. In
particular, we are grateful to members of the Miller labora-
tory, past and present, for their contributions to the ideas
that are presented here. We would also like to apologize to
those authors whose work was not cited owing to space limi-
tations. A.H. is supported by a Career Development Award
from the Northwest Regional Center of Excellence for
Biodefense and Emerging Infectious Diseases Research
(National Institute of Allergy and Infectious Diseases (NIAID)
grant U54 AI057141). M.B.O. is supported by the
Comprehensive Training in Inter-Disciplinary Oral Health
Research T32 grant DE07132. S.I.M. is supported by the
National Institutes of Health, NIAID grants R01 AI30479,
R01 AI048683 and U54 AI057141 for the Northwest
Regional Center of Excellence for Biodefense and Emerging
Infectious Diseases Research.
DATABASES
Entrez Genome Project: http://www.ncbi.nlm.nih.gov/
entrez/query.fcgi?db=geneomeprj
Salmonella typhi | Salmonella typhimurium
All links are active in the online pdfF