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Membrane Vesicles and Horizontal Gene Transfer in
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Membrane vesicles (MVs) are released from all living cells. MVs by secretion from living cells [1,2]. Several models can
are lumen-containing spheres of lipid-bilayers derived from the explain the biogenesis (also called vesiculation) of MVs in
cell surface. MVs are biologically active and contain various processes that are not yet fully understood [2,3]. MVs
components, including genetic material. Both chromosomal are biologically active and a multitude of functions have
and plasmid DNA, as well as different types of RNA have been been associated with these vesicles [1,2,3]. Here we
detected in MVs. Vesicle-mediated transfer of genes coding for review the characteristics of and the role of MVs in
antibiotic resistance, virulence and metabolic traits has been horizontal gene transfer (HGT) processes in prokaryotes.
reported in Gram-negative and Gram-positive bacteria and in
Archaea. MVs can persist over time in natural environments.
Here we review the characteristics of and the role of MVs in
Formation of membrane vesicles (MVs)
horizontal gene transfer (HGT) processes in prokaryotes.
MVs are lumen-containing spheres of lipid-bilayers
derived from the cell surface; occasionally they can be
Addresses elongated or elliptical, especially during vesiculation
1
Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, [4,5]. Their diameter varies and ranges from 10 to
Portugal
2
Center for Neuroscience and Cell Biology, 3004-517 Coimbra, Portugal
500 nm (Table 1). The amount and composition of vesi-
3
Department of Life Sciences and Health, Oslo and Akershus University cles vary even within the same species and population
College, 0130 Oslo, Norway and depends on growth phase and environmental condi-
4
Genok-Center for Biosafety, SIVA Innovation Center, 9294 Troms, tions [1,6,7]. For example, Acinetobacter baumannii
Norway
ATCC19606 populations release small outer MVs in
Corresponding authors: Domingues, Sara (saradomingues@ff.uc.pt), the early log-phase, large MVs in the early and mid-log
Nielsen, Kaare M (kaare.nielsen@hioa.no) phase, and medium sized vesicles called inner and outer
membrane vesicles (IOMVs) comprising elements from
the inner and outer membranes and a putative peptido-
Current Opinion in Microbiology 2017, 38:16
glycan layer, during the stationary phase [8].
This review comes from a themed issue on MGE-HGT in prokaryotes
Edited by Andrew Lang, J Thomas Beatty and Phoebe Rice MVs are secreted in a variety of environments, including
planktonic and biofilm stages, within eukaryotic host
cells, and when present in different growth media in
the laboratory [6,7]. A conserved general mechanism
http://dx.doi.org/10.1016/j.mib.2017.03.012 for the biogenesis of MVs has not been identified so
1369-5274/ 2017 Elsevier Ltd. All rights reserved. far, but several models have been proposed.
Table 2
Figure 1
Phages
DNA
fragments
Recombination
Genome / replication
Free DNA
Donor cell Recipient cell
Membrane vesicles
Current Opinion in Microbiology
Schematic presentation of membrane-vesicle (MV) mediated horizontal gene transfer from a donor to a recipient cell. Membrane vesicles (10
500 nm in diameter) may contain nucleic acids in the lumen or bound to the surface. Upon lysis DNA in the lumen will be sensitive to nucleases.
Bacteriophages may also bind to MVs in the environment leading to transduction-like HGT processes. The process is multi-directional.
and/or of phage origin [7]. Both natural and vector plas- outer surface of MVs [4,15,21,22] or be present in their
mids have been found inside MVs [4,16]. lumen [4,15,2123]. MVs allow transformation in bacte-
rial species that are not considered to efficiently express
Different types of RNA, including mRNA, rRNA, sRNA competence for natural uptake of DNA, such as Escher-
and tRNA, have also been found in MVs (Table 2). In ichia coli and Salmonella enterica. However, in species that
some cases, the RNA present covered the majority of the are efficient at expressing competence for natural DNA
genes of the genome of the MV producing bacterial strain uptake, OMVs-mediated transfer seems to depend also
[19]. on the competence machinery in the recipient cell, at
least in Acinetobacter baylyi [4] and in Thermus thermophilus
Some MVs, such as the ones detected in Neisseria gonor- [27].
rhoeae, Prochlorococcus sp. and Porphyromonas gingivalis,
carried both DNA and RNA [18,21,26]. MVs confer protection of the DNA in the lumen against
nucleases, as seen by the unaffectedness of the DNA
HGT by membrane vesicles content in OMVs [18,26,28] and OMV-mediated trans-
MVs carrying genetic material can upon exposure to new fer after addition of DNase [4,16,29]. MVs have also
host cells result in gene transfer events (Figure 1; been reported to protect DNA from thermodegradation
Table 3). The genetic material can be linked with the [30].
Table 3
The specifics of the mechanism of gene delivery at the However, not all studies of MV transfer have proven
host cell surface often remain unclear. Different pathways successful. Exposure studies with DNA contained inside
seem to occur depending on the recipient species. For OMVs have also failed. For instance, plasmid-containing
instance, OMVs might attach to the OM surface, followed OMVs released by Pseudomonas aeruginosa PAO1 failed to
by lysis; or attach to the OM and then become internal- transform wild-type P. aeruginosa PAO1 and E. coli DH5a
ized by the recipient Gram-negative cell [4]. recipient cells under different growth conditions; failure
was possibly associated with the inability of the plasmid
Successful HGT events require inheritance of the DNA to pass the plasma membrane [15].
acquired genetic material, measured as transmission to
subsequent generations. Expression of genes transferred MV-mediated HGT in other prokaryotes
by MVs has been observed, as well as the heritable nature Fewer studies on the release of MVs are available in
of the HGT events in both Gram-positive [24] and prokaryotes other than Gram-negative bacteria and only
Gram-negative [16,23] bacteria. MVs-mediated gene very few studies have reported the occurrence of MV-
transfer can contribute to the dissemination of microbial mediated HGT. It is unclear if this is the result of lack of
fitness determinants such as antimicrobial resistance [16], studies or publication bias.
metabolic properties [24] and virulence genes [23].
MVs-containing chromosomal DNA released from the
Gram-positive Ruminococcus sp. and Ruminococcus albus
Examples of MV-mediated gene transfer in Gram- were shown to transfer DNA to Ruminococcus sp. recipient
negative bacteria cells, but not to Butyrivibrio fibrisolvens [24].
Fulsundar et al. tracked the route of plasmid DNA from
the bacterial cytoplasm to the lumen of OMVs; the DNA MVs carrying a shuttle plasmid secreted by the hyper-
path was traced by immunogold labelling followed by thermophilic archaea Thermococcus kodakarensis were able
visualization by transmission electron microscopy [4]. In to transform T. kodakaraensis recipient cells, indicating
their laboratory study, OMVs-mediated transfer of plas- that MV-mediated gene transfer also occur in this domain
mid DNA between populations of A. baylyi strains and of life [29].
also to E. coli DH5a recipient cells occurred at frequen-
cies between 10 8 to 10 6 gene transfer event per recipi- We are not aware of studies that have explored the
ent cell; immunogold labelling of the OMVs with FITC possibility of horizontal transfer of RNA mediated by
allowed the observation of attachment and internalization MVs. Horizontal transfer of RNA molecules are rarely
of the vesicles by the recipient cells. A. baylyi were described in bacteria and the biological relevance of such
transformed at a higher frequency with the OMVs than events remain unclear. This is in contrast to the RNA
with naked DNA in the natural transformation assay used. content of mammalian exosomes. Based on the observed
Within the same genus, two clinical strains of A. bauman- effects of transfer of RNA carried by eukaryotic exosomes
nii were also shown to release, during the exponential in new host cells, Sjostrom et al. suggested that similar
growth phase, OMVs carrying plasmid DNA containing a mechanisms can be involved in bacteriabacteria inter-
carbapenemase gene, blaOXA-24. Horizontal transfer of the actions resulting in modulation of expression of target
plasmid to A. baumannii ATCC 17978 recipient cells was genes in recipient cells [20].
seen after OMV exposure and resulted in an increase in
the minimal inhibitory concentrations to several b-lactam Environmental influence on vesiculation and HGT
antibiotics in the recipient strain [16]. Several external stimuli induce the release of MVs,
including temperature, desiccation, starvation, UV light,
Horizontal transfer of plasmid DNA in MVs between N. quorum sensing molecules, oxygen stress and exposure to
gonorrhoeae strains was demonstrated in early studies [26 subinhibitory antibiotics of different classes, such as
]. Chromosomal DNA present in OMVs of P. gingivalis ceftazidime, chloramphenicol, ciprofloxacin, gentamicin
ATCC 49417 was found to transfer to P. gingivalis ATCC and imipenem [4,8,3234]. It can therefore be hypothe-
33277 at a requency of 1.9 10 7. The DNA was inte- sized that HGT rates mediated by OMVs may be
grated into the recipient genome by homologous recom- enhanced by stress. Some stress factors may also increase
bination and subsequently expressed [21]. the overall concentrations of DNA in the cytoplasm.
Hence more DNA may be part of the lumen content
Although not originally called MVs, but virus-like parti- of the MVs [4] under such conditions.
cles, MVs-containing DNA present in natural seawater
were able to transform E. coli with a transformation Discussion
frequency of 1.0 10 2 [25,31]. Yaron et al. showed vesi- Prokaryotes maintain several mechanisms to take up
cles-mediated transfer of plasmid and phage DNA from DNA from the environment and other host cells. MVs
E. coli O157:H7 to E. coli JM109 and phage DNA to S. add to the arsenal of gene transfer mechanisms and
enterica serovar Enteritidis ATCC 13076 [23]. provide a shielded environment for various genetic
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