CHAPTER 37
Intracellular Motility
Virtually every component inside living cells moves to
some extent (Fig. 37.1), but the magnitude and velocity
of these movements vary by orders of magnitude (Table
37.1). At one extreme, the bulk cytoplasm of algae and
giant amoebas streams tens of micrometers per second,
but most cytoplasm is generally less dynamic. The cyto-
plasmic network of cytoskeletal polymers has a pore size
of less than 50nm (see Fig. 1.13). This allows small
molecules and macromolecules to diffuse essentially
unimpaired. Particles larger than the pores must be trans-
ported actively. For example, lysosomes, mitochondria,
secretory vesicles, and endosomes all move actively in
cytoplasm, frequently between the centrosome and the
cell periphery. Similarly, messenger RNA (mRNA) moves
from its site of synthesis in the nucleus through nuclear
pores into the cytoplasm and then may be carried actively
to specific parts of the cell. Intracellular pathogenic bac-
teria and viruses take advantage of the host cells actin
system to propel themselves through the cytoplasm.
Virus particles can also move along microtubules.
Two ancient mechanisms (Fig. 37.1C) account for
most intracellular movements in eukaryotes. Transport
along microtubules by kinesin or dynein predominates
in animal cells. Transport along actin filaments by myosin
is more important in plants and fungi. Specialized iso-
forms of myosin, kinesin, and dynein are dedicated to
particular movements. Experiments with drugs that
depolymerize or stabilize actin filaments or microtubules
(see Boxes 33.1 and 34.1) originally identified the cyto-
skeletal polymers that support various biological move-
ments. Discovering the motors was more challenging,
given multiple genes for myosin, kinesin, and dynein in
most organisms and a limited choice of pharmacologic
agents (Box 37.1). Even loss of function mutations and
depletion by RNA interference (RNAi; see Fig. 11.12)
have limitations, because some motors are essential for
viability while others contribute redundantly to trans-
port. For example, dynein contributes to mitosis, but
Drosophila tissue culture cells depleted of dynein can
complete mitosis, because other motors take over allow-
ing mitosis to proceed, albeit more slowly.
The diversity of cargo, motors, and tracks poses a
traffic control challenge, which cells manage by specify-
ing the organization of microtubules and actin filaments,
matching appropriate motors to specific cargo and regu-
lating motor activity. Most cargos associate with more
than one type of motor, so their activities must be coor-
dinated. For example, some cargos with two motors
reverse their direction either along the way or after
reaching their destination. Local or global signaling path-
ways influence many of these choices, allowing cells to
adjust transport to adapt to environmental conditions.
This chapter takes a broad view across biology, high-
lighting the wide range of mechanisms that produce
intracellular transport. In most cases, transport involves
single organelles on an actin filament or microtubule,
but organelle transport can result in bulk movement of
the cytoplasm. The chapter also considers mechanisms
that produce special types of intracellular movements:
cytoplasmic contractions generated by myosin and
actin filaments that propel cytoplasmic streaming; and
polymerization and depolymerization of microtubules
and actin filaments. Chapters on membrane traffic (see
Chapters 20 to 22) and mitosis (see Chapter 44) cover
more examples of intracellular movements.
Rapid Intracellular Movements
on Microtubules
Organelles in most eukaryotic cells can move along
linear microtubule tracks at relatively high velocities, on
the order of 1ms1 (Table 37.1) at least part of the
time. The physical organization of microtubules deter-
mines the patterns of these movements (Fig. 37.1; also
see Fig. 34.2). In cells with a radial arrangement of micro-
tubules (Fig. 37.1) organelles and other cargos associated
with kinesin-1 move toward microtubule plus ends
located at the periphery of cells. Cargo associated with
639
640 SECTION IX n Cytoskeleton and Cellular Motility

A B

Nitella streaming

Neuron

Axon
Fibroblast

Synapse
Myosin
Kinesin

Dynein

FIGURE 37.1 MECHANISMS OF INTRACELLULAR MOVEMENT. A, Fibroblasts and neurons move organelles bidirectionally along micro-
tubules (red). B, The green alga Nitella moves cytoplasmic organelles along bundles of actin filaments located in the cell cortex.
C, Microtubule-based and actin filamentbased motors.

TABLE 37.1 Velocities of Intracellular Movements

System Velocity (m s1) Mechanism
Microtubule Motors
Anterograde fast axonal transport, squid 1 Individual kinesin motors
Retrograde fast axonal transport, squid 2 Individual dynein motors
Chromosome movement in anaphase of mitosis 0.0030.2 Motors plus depolymerization
Endoplasmic reticulum sliding, newt cell 0.1 Individual kinesin motors
Slow axonal transport, rat nerves 0.0020.1 net 1 (intermittent) Motors on microtubules
Microtubule Polymerization
Endoplasmic reticulum tip elongation, Newt cell 0.1 Microtubule polymerization
Actin-Myosin Motors
Cytoplasmic streaming, Nitella 60 Myosin motors on tracks
Cytoplasmic streaming, Physarum 500 Actinmyosin contraction
Actin Polymerization
Actin-propelled comet, Listeria 0.5 Actin polymerization

dynein moves in the opposite direction. These motors
are processive (see Chapter 36), so they can work alone
or in small numbers. Intraflagellar transport of proteins
in cilia and flagella (see Fig. 38.18) shares many features
with the movements of organelles.
The chemistry of individual microtubules subtly influ-
ences the delivery of cargo to specific locations, as
some motors favor microtubules composed of particular
tubulin isoforms or with posttranslational modifications.
For example, kinesin-1 favors acetylated microtubules,
and CENP-E favors tyrosinated microtubules in the center
of the mitotic spindle. In addition, microtubule-associated
proteins, such as tau, can influence the choice of tracks
by reducing the run lengths of motors under some
circumstances.
Matching Microtubule Motors With Cargo
Pairing motors with appropriate cargo and regulation of
motor activity control the traffic along microtubules.
Coupling can be as simple as kinesin binding to a
 CHAPTER 37 n Intracellular Motility 641

A p25/p27 B C
Cargo Dynactin
p62 Lysosome
Pointed
end Arl8-GTP
BICD2
Arp11 SKIP
RU
-actin Dynein N
(ADP.Pi)
WD PH
Arp1
Dyenin KLC
Kinesin-1
Arp1 (ADP)
Filament Shoulder Anterograde
p150Glued/p135 (+) movement
p50, p24
()

Bottom protofilament
Microtubule
Barbed Top protofilament
end (+)
Capping protein ()

FIGURE 37.2 ATTACHMENT OF CYTOPLASMIC MICROTUBULE MOTORS TO MEMBRANES. A, Ribbon diagram of the structure of
the core of the dynactin complex determined by electron cryomicroscopy. The short filament of Arp1 is capped by capping protein on the barbed
end and by Arp11 on the pointed end. The shoulder is formed by part of the p150 molecule, most of which was disordered in these samples.
B, Drawing showing dynein linked to a vesicle by dynactin complex and moving toward the minus end of a microtubule. C, Coupling of kinesin-1
to a lysosome. Guanosine triphosphatase (GTPase) Arl8 on the lysosome membrane engages the adapter protein SKIP (SifA-kinesin interacting
protein), which binds to the kinesin light chain as shown in Fig. 36.9E. (A, From Urnavicius L, Zhang K, Diamant AG, etal. The structure of the
dynactin complex and its interaction with dynein. Science. 2015;347:14411446. B, From Cianfrocco MA, DeSantis ME, Leschziner AE, Reck-
Peterson SL. Mechanism and regulation of cytoplasmic dynein. Annu Rev Cell Dev Biol. 2015;31:83108. C, Based on a drawing from Rosa-
Ferreira C, Munro S. Arl8 and SKIP act together to link lysosomes to kinesin-1. Dev Cell. 2011;21:11711178.)

Other subunits at the pointed end serve as cargo adap-

BOX 37.1 Tools for Studying Motor Proteins
A small compound named monastrol inhibits kinesin-5,
resulting in monopolar mitotic spindles that fail to
segregate the chromosomes. Higher-affinity inhibitors of
kinesin-5 are being tested for cancer therapy. The small
molecule blebbistatin inhibits cytoplasmic and skeletal
muscle myosin-II (but not smooth muscle myosin-II) and
blocks cytokinesis. Vanadate and ultraviolet light can inac-
tivate dynein. Vanadate binds to the -phosphate site of
dyneinadenosine diphosphate (ADP), but it binds simi-
larly to other adenosine triphosphatases (ATPases), so it is
not specific. However, ultraviolet light has a novel effect
on the dyneinADPvanadate complex: It cleaves and
inactivates the dynein heavy chain.
transmembrane protein on a cargo vesicle, but usually
involves adapter proteins that recognize a molecule on
the surface of the cargo and one or more motors. Dynein
associates with the dynactin complex (Fig. 37.2B).
About a dozen other known adapter proteins also
interact with the dynactin complex, kinesins or both to
direct traffic to specific locations (Table 37.2). Local
conditions in the cell regulate both coupling of motors
to cargo and the activity of the motors. Other proteins
tether some organelles to immobilize them and prevent
their transport.
The 1mDa dynactin complex consists of a short fila-
ment of actin-related protein Arp1 capped on its barbed
end by capping protein and on its pointed end by the
most divergent actin-related protein, Arp11 (Fig. 37.2A).
tors. Association of the dimeric tail of dynein with the
dynactin complex and adapter proteins activates its
motor activity. The 150-kD dynactin subunit (p150Glued)
includes a long extension that links the Arp1 filament to
both an intermediate chain of dynein and a microtubule.
The two dynein motor domains take steps of variable
size as they pull the cargo along the microtubule. Muta-
tions of p150Glued cause developmental defects in the eye
and brain of Drosophila plus some forms of Parkinson
disease and motor neuron degeneration in humans.
Most proteins known to link organelles and ribonu-
cleoprotein particles to microtubule motors interact
with both dynein and kinesins, but some are specific for
one or the other (Table 37.2). These interactions often
activate or inhibit motor activity, coupling physical
association with transport. For example, some cargo
adapters overcome autoinhibitory interactions between
the heads and tails of kinesins-1, -2, and -3 by binding to
the kinesin tail or light chains.
Other than their linker and regulatory activities,
adapter proteins have little in common, so they likely
had independent evolutionary origins. The following
sample illustrates the diversity of these adapters.
Motors and adapter proteins are targeted to cargo
membranes by integral membrane proteins, peripheral
proteins or lipids. Kinesin binds directly to a few trans-
membrane proteins, so no adapter is required. For
example, calsyntenin and alcalpha on certain neuronal
vesicles have short sequences with acidic residues
flanking a key tryptophan that bind kinesin light chains
642 SECTION IX n Cytoskeleton and Cellular Motility

TABLE 37.2 Adapters for Microtubule Motors

Disease
Adapter Protein Links to Motors Links to Cargos Cargo Destination Regulation Connection
Dynein Only
Dynactin Dynein IC, Endosomes, Mutations
complex p150Glued, other mitochondria, cause neuron
adapters listed Golgi membranes degeneration
here
RILP (Rab7- Dynactin Rab7, spectrin Late endosomes Trans-Golgi Rab7 GTPase
interacting network
lysosomal
protein)
SNX6 (sorting Dynactin Other retromer Endosome vesicles Trans-Golgi PI4P on trans-
nexin 6) proteins with recycling network Golgi
receptors membranes
releases dynactin
Kinesin Only
DENN/MADD Kinesin-3 Synaptic vesicles Synapse Rab3 GTPase
(Rab3 guanine with Rab3
nucleotide
exchange
factor)
Mint1 (Munc18- Kinesin-2 tail Vesicles with Synapse Phosphorylation
interacting NMDA glutamate by CaMKII
protein) receptors
SKIP (SifA-kinesin Kinesin-1 LC Arl8 GTP Lysosomes Periphery Arl8 GTPase
interacting lysosome
protein)
Dynein and Kinesin
Hook Dynein, kinesin-3 Early endosomes Bidirectional
Huntingtin Dynein IC, HAP1 HAP40 Rab5, Endosomes, Bidirectional Phosphorylation Traffic defects
dynactin, Optineurin lysosomes, of Huntingtin and massive
kinesin-1 HC myosin-VI autophagosomes, and motors neuronal
and LC vesicles with degeneration
APP or GABA in Huntington
receptors, mRNA disease
JIP1 (JNK Either dynactin or JNK, vesicles Synaptic vesicles, Bidirectional Phosphorylation
interacting kinesin-1 stalk, with amyloid autophagosomes, by JNK favors
protein) tail & LC with precursor mitochondria, kinesin-1 binding
aid of FEZ1 protein vesicles with APP and transport to
plus end
JIP3/JIP4 (JNK Dynactin, JNK, Arf6 Synaptic vesicles Phosphorylation
interacting kinesin-1 tail to synapse, of DIC
protein) and LC, JIP1 endosomes with
nerve growth
factor to cell
body
La Dynein IC, RNPs Ribonucleoprotein Bidirectional SUMOylation may
kinesin-1 HC particles reverse direction
at periphery
Milton/TRAK Dynactin, Miro/RhoT2 Mitochondria Bidirectional Local Parkinson
kinesin-1 HC transmembrane concentrations disease (?)
GTPase of ATP and Ca2+

Data from Fu MM, Holzbaur EL: Integrated regulation of motor-driven organelle transport by scaffolding proteins. Trends Cell Biol 2014;24:564574.
APP, amyloid precursor protein; ATP, adenosine triphosphate; CaMKII, calcium/calmodulin-dependent serine protein kinase; FEZ1, fasciculation
and elongation protein zeta 1; GABA, -aminobutyric acid; GTP, guanosine triphosphate; GTPase, guanosine triphosphatase; HC, heavy chain;
IC, intermediate chain; JNK, c-Jun N-terminal kinase; LC, light chain; mRNA, messenger RNA; NMDA, N-methyl-D-aspartate; RNP, ribonucleoprotein;
PI4P, phosphatidylinositol 4-phosphate.
 CHAPTER 37 n Intracellular Motility 643

(see Fig. 36.9E). In other cases an adapter protein links
a transmembrane protein to a motor. For instance, a
transmembrane protein called Miro on the surface of
mitochondria interacts with adapter protein Milton/
TRAK1, which binds both the kinesin-1 heavy chain and
dynactin for bidirectional transport in axons.
Guanosine triphosphatases (GTPases) on organelle
membranes interact with other adapter proteins. For
instance, different GTPases target kinesin and dynein
to lysosomes. Lysosomes with Arl8-GTP (guanosine tri-
phosphate) bind the adapter SKIP (SifA-kinesin inter-
acting protein). An acidic/tryptophan sequence in SKIP
interacts with kinesin light chains (Fig. 37.2C) during
transport to the periphery. This interaction with SKIP
also turns on the kinesin motor by overcoming the auto-
inhibitory interaction of the motor domains with the
tail. Similarly, vesicles containing transmembrane recep-
tors (mannose-6-phosphate receptor, for example) bud
from late endosomes with the GTPase Rab7 on their
surface. Rab7-GTP binds the adapter protein RILP
(Rab7-interacting lysosomal protein), which anchors
dynactin and dynein for vesicle transport to the trans-
Golgi network (see Fig. 22.12). Phosphatidylinositol
4-phosphate (see Fig. 26.7) on the Golgi membranes
binds sorting nexin 6 (part of the retromer complex; see
Fig. 22.12) and releases dynactin and dynein.
Many cargo particles associate with both a kinesin and
dynein. In this case, regulatory mechanisms responsive
to local conditions must coordinate their activities to
achieve net transport. One example is the adapter protein
La, which links ribonucleoprotein particles to both
dynein and kinesin. After being carried to the cell periph-
ery by kinesin, covalent modification of La with a SUMO
protein inactivates kinesin, allowing for reverse transport
back to the cell center by dynein. Phosphorylation regu-
lates the adapter protein JIP1 (JNK interacting protein-1),
which binds either kinesin or dynein. Phosphorylated
JIP1 binds and activates kinesin, favoring anterograde
transport. Mitochondria move both directions in axons,
but some stop moving in regions with limited adenosine
triphosphate (ATP). A local high concentration of cyto-
plasmic calcium binds to EF-hands (Ca2+ binding site in
calmodulin consisting of -helices E and F) of the anchor
protein Miro. Calcium binding turns off both kinesin and
dynein, keeping mitochondria in place until ATP concen-
trations return to normal. Another protein anchors sta-
tionary mitochondria to microtubules.
Faulty axonal transport resulting in clumps of vesicles
is observed in human degenerative diseases of the
nervous system including Alzheimer and Parkinson dis-
eases, but cause-and-effect relationships are still under
investigation. One intriguing part of this story is that
the transmembrane amyloid precursor protein not only
binds directly to kinesin-1 light chain, but also is cleaved
in Alzheimer disease to produce amyloid- peptidea
toxic peptide that is implicated in neuronal death. Hun-
tingtin, the giant (350kD) adapter protein mutated in
Huntington disease, normally participates in bidirec-
tional axonal transport of multiple cargos (Table 37.2).
Other membrane-associated scaffold proteins, including,
JIP-1 and JIP-3, bind kinases from the mitogen-activated
protein (MAP) kinase cascade (see Fig. 27.6) in addition
to kinesin-1 light chains.
Fast Axonal Transport
Analysis of microtubule-based movements is particularly
favorable in axons of neurons, because axons are long
(up to 1m) but narrow, the microtubules have a uniform
polarity, and organelles move at steady rates in both
directions. Furthermore, nerve cells contain high con-
centrations of microtubules and microtubule motors;
indeed, cytoplasmic tubulin, cytoplasmic dynein, and
kinesin were all originally isolated from brain.
High-contrast light microscopy of living axons reveals
that most membrane-bound organelles move either
toward (anterograde) or away from (retrograde) the
end of the axon (Fig. 37.3) with some pauses and even

0 sec 3 sec 5 sec

2 1 2
2

1 1

4
4 4
A 3 3 3 B
FIGURE 37.3 FAST TRANSPORT IN CYTOPLASM ISOLATED FROM SQUID GIANT AXONS. A, Three frames from a series of video-
enhanced differential interference contrast micrographs show movement of organelles in both the anterograde (rightwards) and retrograde (left-
wards) directions. Four large organelles are marked with numbers and colored green at zero time, blue at 3 seconds, and red at 5 seconds.
Movement (arrows in right panel) is from the white to the black number. The original video record shows hundreds of smaller organelles moving
steadily in either an anterograde or a retrograde direction at 1 to 2m/s. B, Electron micrograph of a thin section showing vesicles associated
with microtubules in axoplasm. (A, Courtesy S. Brady, University of Texas Southwestern Medical School, Dallas. B, Courtesy R.H. Miller, Case
Western Reserve Medical School, Cleveland, OH.)
644 SECTION IX n Cytoskeleton and Cellular Motility

A B
Proximal Ligature Distal
FIGURE 37.4 ELECTRON MICROGRAPHS SHOWING THE RESULT OF NERVE LIGATION. These are longitudinal sections of axons
surrounded by a darkly stained myelin sheath. A, The cytoplasm proximal to the ligation demonstrates the accumulation of vesicles and mito-
chondria, which were being transported toward the nerve terminal to the right. B, The cytoplasm distal to the ligation shows the accumulation
of lysosomes, multivesicular bodies, and mitochondria, which were being transported toward the cell body to the left. (B, From Hirokawa N,
Sato-Yoshitake R, Yoshida T, etal. Brain dynein (MAP1C) localizes on both anterogradely and retrogradely transported membranous organelles
in vivo. J Cell Biol. 1990;111:10271037.)

occasional changes of direction. Retrograde move-
ments (2.5ms1 or 22cm/day) are faster than antero-
grade movements (0.5ms1 or 4cm/day). These
rates are remarkable given the densely packed microtu-
bules, intermediate filaments, and vesicles in the cyto-
plasm (Fig. 37.4; see also Fig. 35.9). At these rates, a
round trip from a nerve cell body in the spinal cord of
a human to the foot and back takes only 3 weeks. If a
0.1-m vesicle were the size of a small car, it would move
anterogradely at 50 miles per hour and retrogradely at
250 miles per hour. In the axons of vertebrate neurons,
mitochondria move back and forth in both directions.
Their net movement toward the nerve terminal or cell
body depends on physiological conditions.
Biochemical reconstitution showed that the plus-end
motors of the kinesin family are responsible for antero-
grade movements toward the nerve terminal and the
minus-end motor dynein is responsible for movement in
the retrograde direction. Accordingly, kinesin mutations
in flies result in paralysis of the back half of larvae,
because transport fails in the longest axons. Mutations
in three different kinesin genes also cause human nerve
degeneration.
Classic nerve ligation experiments revealed the cargo
carried in each direction by fast transport (Fig. 37.4).
Different organelles pile up on either side of a mechani-
cal constriction that blocks transport. Small, round, and
tubular vesicles, including components of synaptic vesi-
cles (see Fig. 17.8), accumulate on the side near the cell
body. Fast anterograde transport carries these cargoes
from the cell body toward the end of the axon, where
they enter the cycle of synaptic vesicle turnover (see
Figs. 17.8 and 17.9). Endosomes and multivesicular
bodies moving by fast retrograde transport pile up on
the distal side of the constriction. Autophagic vesicles
also move primarily in the retrograde direction. Retro-
grade transport can also move signals from nerve termi-
nals to the cell body. For example, kinesin carries vesicles
with the nerve growth factor TrkA receptor tyrosine
kinase (see Fig. 24.4) to nerve terminals where it joins
the plasma membrane. When activated by nerve growth
factor, TrkA is taken up by endocytosis for transport by
dynein back to the perinuclear region. Once the vesicle
reaches the cell body TrkA activates the MAP kinase
pathway to regulate cell growth (see Fig. 27.6). Herpes
virus and rabies virus also use dynein for long-distance
transport on microtubules from the terminals of sensory
nerves to the cell body, where viral DNA enters the
nucleus for replication.
Many questions remain regarding the control of
microtubule motors during fast transport, including how
kinesins remain active and dynein remains inactive
during the long trip out to the nerve terminal, how cyto-
plasmic dynein on retrograde cargo is activated locally
at the nerve terminal and kept active during movement
to the cell body, and how defects in fast transport may
contribute to neurodegenerative diseases.
Slow Transport of Cytoskeletal Polymers
and Associated Proteins in Axons
Many neuronal proteins move slowly from their site of
synthesis in the cell body toward the ends of axons and
dendrites. This transport is essential, as most protein
 CHAPTER 37 n Intracellular Motility 645

A. Radiolabeling pulse-chase protocol B. Photobleaching C. Fluorescence microscopy of a

neurofilament moving through
Inject radiolabeled amino acids into eye Inject fluorescently a bleached zone in an axon
where neurons synthesize them into proteins labeled tubulin
Anterograde

0
Bleached zone

56

64
Uniformly fluorescent
axon

Time (sec)
Photobleach 72
discrete zone
Wave of labeled proteins
moves along axons
toward brain 80
Bleached zone

88

96
Wave of labeled proteins
progresses 1-2 mm/day Zone remains
stationary

FIGURE 37.5 EXPERIMENTS ON SLOW AXONAL TRANSPORT. A, Pulse-chase experiment. Radioactive amino acids are injected into
the eye of an experimental animal. In the nerve cell body, radioactive tracer is incorporated into proteins, which are transported along the axon.
Some proteins are incorporated into stationary structures and are left along the way. B, Photobleaching experiment. A cultured nerve cell is
injected with tubulin labeled with a fluorescent dye. Tubulin fills the cytoplasm and axon as it grows out. A section of the axon is then bleached
with a strong pulse of light. This bleached zone is stationary over a period of minutes. C, Fluorescence micrographs of the axon of a cultured rat
neuron showing rapid transport of a short neurofilament labeled with subunits fused to green fluorescence protein (GFP). Note the photobleached
region (bracket) and the ends of the moving neurofilament (arrows). The neurofilament moves rapidly into the bleached region, but the bleached
region does not move because most of the neurofilaments are stationary. Scale bar is 5m. (AB, Modified from Cleveland DW, Hoffman PN.
Slow axonal transport models come full circle. Cell. 1991;67:453456. C, From Wang L, Brown A. Rapid intermittent movement of axonal neu-
rofilaments observed by fluorescence photobleaching. Mol Biol Cell. 2001;12:32573267, 2001.)

synthesis occurs in the cell body, whereas more than
99% of cell volume can be in axons and dendrites. If
nerve cells were smaller, shorter lived or less asymmetri-
cal, we might not even notice such slow movements.
Labeling proteins with radioactive amino acids during
their synthesis in the cell body allows for tracking their
movements along axons (Fig. 37.5A). Proteins that are
moved by slow axonal transport are classified into two
groups based on their velocities. Tubulin, intermediate
filament proteins, and spectrin, which compose the
slow componenta, move exceedingly slowly, approx-
imately 0.1 to 1.0mm per day (or 1 to 10nms1). In a
human, these molecules take more than 3 months to
travel from their site of synthesis in the spinal cord to
the foot. Slow componentb moves approximately 10
times faster and includes 10 times more protein than
slow componenta. It is a heterogeneous mixture of
many proteins, including clathrin, glycolytic enzymes,
and actin.
Defining the mechanism of slow transport was
challenging because various experimental approaches
yielded apparently conflicting results. Radioactive label-
ing showed that the moving proteins are spread out
and diluted as they move away from the cell body
(Fig. 37.5A). Photobleaching of fluorescent tubulin and
actin in axons of cultured neurons demonstrated that
the bulk of those cytoskeletal polymers is stationary
(Fig. 37.5B).
This puzzle was solved for slow componenta by
imaging single fluorescent intermediate filaments in
axons of live nerve cells. These filaments are stationary
most of the time (up to 99%), but occasionally, they
move rapidly (0.2 to 2ms1) for up to 20m. Most of
these intermittent movements are away from the cell
646 SECTION IX n Cytoskeleton and Cellular Motility

body, accounting for the net anterograde movement.

A B C
These rapid but intermittent movements depend on
microtubules and are presumably driven by kinesin,
although how the motor is linked to the filaments is not 00:00 00:00
known. A few intermediate filaments move in the retro-
grade direction explaining why waves spread in pulse
chase experiments (Fig. 37.5A). The movements of
whole microtubules are similar to those of intermediate 00:35 01:00
filaments. Thus, what appears to be slow bulk transport 00:00
is caused by fast but intermittent movement of cargos.
Slow componentb proteins seem move by alternatively
hitchhiking transiently on some moving structures and 00:42 02:00
diffusing locally between movements.

Other Microtubule-Dependent Movements

00:49 02:10
Other cells use the same molecular mechanisms as
neurons to move organelles in the cytoplasm. Secretory 06:24
vesicles use plusend motors to move from the Golgi
apparatus to the plasma membrane (see Fig. 21.1). Endo- 02:06 02:49
somes use dynein to move from the plasma membrane
toward the centrosome.
The intracellular distributions of the endoplasmic
reticulum (ER) depend on microtubules. Strands of the 03:02 03:30
ER align with microtubules in cultured cells (Fig. 37.6).
This codistribution is achieved in two ways: (a) Motors
transport strands of ER bidirectionally on microtubules, 12:48
and (b) other strands of the ER attach to the plus end of 03:30 04:00
microtubules and ride the microtubule tip as it grows
and shrinks during dynamic instability. The +TIP EB1
(see Fig. 34.8) interacts with transmembrane protein
STIM1 to couple the ER to growing microtubules. STIM1 03:58 04:10
has another role in maintaining a supply of Ca2+ inside
the ER (see Fig. 26.12). This is the best example of move-
ment of an organelle driven by forces generated directly
by microtubule assembly. 19:15 04:40 04:40
Microtubules and motors also distribute other organ-
FIGURE 37.6 TWO MODES OF MICROTUBULE-DEPENDENT
elles inside cells. The concentration of the Golgi appara- MOVEMENT OF THE ENDOPLASMIC RETICULUM IN A NEWT
tus near the centrosome depends on microtubules, EPITHELIAL CELL. The cell was microinjected with rhodamine-
because dynein motors transport Golgi vesicles toward labeled tubulin, which incorporates into microtubules, and a lipophilic
the minus ends of the microtubules nucleated at centro- green-fluorescent dye (DiOC6 [3,3-dihexyloxacarbocyanine iodide]),
which labels endoplasmic reticulum (ER). Time series are indicated
somes (see Fig. 21.20). Mitochondria move bidirection-
in minutes and seconds. Scale bar for all panels is 5 M. A, This
ally on microtubules in animal cells but depend on actin column of fluorescence micrographs illustrates the dynamics of
filaments in yeast. Thus, not only the dynamics of the microtubules (red) and ER (green), over a period of 19 minutes.
organelles but also the overall organization of a cell Note the strand of ER moving away from the leading edge (arrow-
depend on microtubules and associated motors. As a heads). B, Time course of the movement of a strand of ER toward
the end of a microtubule, followed by retraction. This type of move-
result, cellular architecture is determined actively, not
ment is thought to be driven by a kinesin motor attached to the
passively. tip of the elongating membrane (arrowhead). C, Time course of the
The intracellular distribution of nucleoprotein com- movement of a strand of ER attached to the tip of a growing
plexes also depends on active movements. The most microtubule (arrowhead), followed by retraction of the membrane
obvious example is the movement of chromosomes along the microtubule. (Courtesy C. Waterman-Storer and E.D.
Salmon, University of North Carolina, Chapel Hill. For reference, see
during mitosis, which depends on microtubule assembly
Waterman-Storer C, Salmon ED. Endoplasmic reticulum membrane
and microtubule motors (see Fig. 44.7). Microtubules tubules. Curr Biol. 1998;8:798806.)
and motors can also produce asymmetrical distributions
of mRNAs in cells. The adapter protein La links kinesins
and dynein to specific ribonucleoprotein particles for

() Nucleus
RNA granules
Disperse
RNA
Branch
Membrane
(+)
Immobile granules
Mobile granules
Disperse RNA
FIGURE 37.7 TRANSPORT OF MESSENGER RNA (mRNA)
FOR MYELIN BASIC PROTEIN IN A CULTURED OLIGODEN-
DROCYTE, A GLIAL CELL ISOLATED FROM BRAIN. mRNA syn-
thesized in the cell body (or, in this case, labeled with a fluorescent
dye and microinjected into the cell body) is packaged with proteins in
a ribonucleoprotein particle, transported from the cell center along
microtubules at a steady rate of 0.2ms1, and released at the
periphery, where it moves randomly at 1m/s. (Modified from Ainger
K, Avossa D, Morgan F, etal. Transport and localization of exogenous
myelin basic protein mRNA microinjected into oligodendrocytes. J Cell
Biol. 1993;123:431441.)
long distance transport to sites of local protein synthesis
(Fig. 37.7). For example, localization of certain mRNAs
in fly oocytes helps establish the polarity of the embryos.
Intracellular Movements Driven by
Microtubule Polymerization
Microtubule polymerization and depolymerization have
long been known to play a central role in the assembly
of the mitotic apparatus and the movement of chromo-
somes (see Fig. 44.7), as well as the establishment of
cellular asymmetry (see Fig. 34.2). Polymerizing micro-
tubules can exert substantial forces, but strong forces
buckle microtubules longer than approximately 10m.
Consequently, microtubule pushing mechanisms work
best over short distances, such as positioning the ER
(Fig. 37.6), the nucleus in fission yeast cells (see Fig.
6.3D) and the mitotic spindle in budding yeast cells
(see Fig. 34.2D).
Remarkably, the end of a depolymerizing microtu-
bule can also pull on attached cargo. An in vitro proof-
of-principle experiment (Fig. 37.8) showed that
chromosomes could ride on the end of a depolymerizing
microtubule, even in the absence of ATP or GTP. Linker
proteins associated with the kinetochore region of the
CHAPTER 37 n Intracellular Motility 647
0 42 51 63 75
FIGURE 37.8 TRANSPORT OF AN ISOLATED CHROMOSOME
ON A SHORTENING MICROTUBULE IN VITRO. A microtubule was
grown from brain tubulin nucleated by a basal body in the extracted
carcass of a ciliate, Tetrahymena. A chromosome (arrow) was added
as a test cargo and captured by the end of the microtubule. When the
concentration of tubulin was reduced, the microtubule shortened, car-
rying along the chromosome attached to its tip. This transport occurs
in the absence of adenosine triphosphate (ATP) or guanosine triphos-
phate (GTP). (Courtesy J.R. McIntosh, University of Colorado, Boulder.)
chromosome make multiple weak bonds with the walls
of the microtubule end (see Figs. 8.21 and 8.22). These
interactions rearrange rapidly enough to maintain attach-
ment, even as tubulin subunits dissociate from the end.
Bulk Movement of Cytoplasm Driven by
Actin and Myosin
Bulk streaming of cytoplasm is most spectacular in plant
cells (Fig. 37.1B). Although confined within rigid walls,
plant cell cytoplasm streams vigorously at very high
velocities (up to 60ms1). At this rate, cytoplasm
moves 5m/day! Such cytoplasmic streaming is best
understood in the giant cells of the green alga Nitella.
Streaming occurs continuously in a thin layer of cyto-
plasm between the large central vacuole and chloro-
plasts immobilized in the cortex. On each side of the
cell, a zone of stationary cytoplasm separates streams
moving in opposite directions. The physiological
function of this streaming is not clearly understood.
Bulk streaming in Nitella is brought about by move-
ment of ER along tracks consisting of bundles of polar-
ized actin filaments associated with chloroplasts (Fig.
37.9C). All the actin filaments in these bundles have the
same polarity, and cytoplasm streams toward their
barbed ends. In Nitella extracts, membrane vesicles
move along actin filament bundles at the same high
velocities as the cytoplasmic streaming. A type XI myosin
(see Fig. 36.7) pulls the ER along these cortical actin
tracks, dragging along other cytoplasmic components,
including organelles and soluble molecules. This myosin
moves nearly 10 times faster than the fastest muscle
contraction, apparently by taking large steps rapidly and
by the cooperation of several motors working rapidly on
the same membrane.
A completely different actomyosin mechanism pro-
duces equally spectacular cytoplasmic streaming in the
acellular slime mold Physarum. In these giant, multi-
nucleated cells, cytoplasm flows back and forth rhyth-
mically at high velocities through tubular channels
(Fig. 37.10). Cycles of contraction and relaxation of
648 SECTION IX n Cytoskeleton and Cellular Motility

A B

C E F
FIGURE 37.9 CYTOPLASMIC STREAMING IN THE GREEN ALGA NITELLA. A, A pair of differential interference contrast (DIC) light
micrographs showing the movement of organelles in cytoplasm. Note the strand of endoplasmic reticulum (ER [arrow]). B, Time series of DIC
light micrographs showing movement of a vesicle isolated from Nitella along a bundle of actin filaments isolated from Nitella. C, Scanning electron
micrographs of the cortex isolated from Nitella showing the bundles of actin filaments associated with chloroplasts. D, Transmission electron
micrographs of a freeze-fracture preparation (upper) and thin section (lower) showing ER associated with actin filament bundles. E, Freeze-fracture
preparation of a vesicle associated with an actin filament bundle. F, Movement of ER along actin filament bundles dragging along bulk cytoplasm.
(Courtesy B. Kachar, National Institutes of Health. For reference, see Kachar B, Reese T. The mechanism of cytoplasmic streaming in characean
algal cells: sliding of endoplasmic reticulum along actin filaments. J Cell Biol. 1988;106:15451552.)

cortical actin filament networks push relatively fluid
endoplasm back and forth in a manner akin to squeezing
a toothpaste tube. Myosin-II is thought to generate the
cortical contraction, as it is present in high concentra-
tion in this cell and can contract actin filament gels in
vitro. (This was the first nonmuscle myosin to be purified
in the late 1960s.) Cortical contractions similar to those
of Physarum are used by giant amoebas for cell locomo-
tion (see Fig. 38.1), and by other cells for cytokinesis
(see Fig. 44.23), and movements of some embryonic
tissues (see Fig. 38.5).
Actin-Based Movements of Organelles in
Other Cells
Like Nitella, budding yeast cells transport vesicles along
bundles of actin filaments from the mother to the bud
(Fig. 37.11), although the movements of these solitary
vesicles do not produce cytoplasmic streaming. Myosin-V
is the motor (see Fig. 36.7), so vesicles fail to move from
mother to bud in null mutants of myosin-V genes.
Myosin-V also transports certain mRNAs along actin fila-
ment cables from the mother to the bud, where they
determine cell fate. Similarly myosin-VII and myosin-X
transport cytoplasmic and membrane proteins in filopo-
dia of animal cells.
Animal cells generally use extended microtubules for
long-distance movements and shorter actin filaments for
local transport of vesicles and RNAs. Fish skin cells use
both systems to change color: dynein aggregates and
kinesin disperses pigment granules called melanophores
along radial microtubule tracks. Myosin-V contributes
by moving dispersed melanophores laterally between
microtubules.

M
5 cm
A B
24
Balance-pressure (cm of water)
20
16
12
8 messenger RNAs (mRNAs), and at least one enzyme (chitin synthase)
4
0
-4 5 10
-8
Time (min)
-12
C
FIGURE 37.10 CYTOPLASMIC STREAMING IN THE ACEL-
LULAR SLIME MOLD PHYSARUM POLYCEPHALUM. A, Photo-
graph of Physarum polycephalum, a giant multinucleated single cell
growing in a baking dish. B, Blur photomicrograph made with polariza-
tion optics by taking a time exposure showing the bulk streaming of
the endoplasm in a cytoplasmic strand (long arrow). M, Mucus.
C, Time course of pressure changes produced by shuttle streaming
of cytoplasm through a strand. (B, From Nakajima H. The mechano-
chemical system behind streaming in Physarum. In Allen RD, Kamiya
N, eds. Primitive Motile Systems in Cell Biology. New York: Academic
Press; 1964:111123. C, For reference, see Kamiya N. The mecha-
nism of cytoplasmic movement in a myxomycete plasmodium. Symp
Soc Exp Biol. 1968;22:199214.)
The direction of movement depends on the motor
and the organization of the actin filaments. Although
many actin filaments in animal cells are not uniformly
polarized, those in the cortex tend to have their barbed
ends near the plasma membrane, allowing for local direc-
tional transport. For example, myosin-V moves recycling
endosomes toward the plasma membrane. Similarly,
myosin-V transports pigment granules called melano-
somes within and between cells in the skin. In both cases
a small GTPase and an adapter protein link melanosomes
to the tail of myosin-V. Mutations of myosin-V, the
GTPase, or the adapter protein in mice and humans
cause not only pigmentation defects but also neurologi-
cal problems owning to faulty mRNA transport in
neurons. Other adapter proteins link myosin-VI to newly
internalized endosomes with various types of receptors
for transport away from the plasma membrane.
CHAPTER 37 n Intracellular Motility 649
A B
C D
FIGURE 37.11 Fluorescence micrographs (AD) showing actin fila-
ment bundles and patches at various stages in the cell cycle of the
budding yeast Saccharomyces cerevisiae. Myosin-V uses these actin
filament bundles to deliver vesicles (including the vacuole), certain
from the mother to the bud. (Courtesy J.A. Cooper, Washington
University, St. Louis, MO.)
Movements Driven by
Actin Polymerization
Some intracellular pathogenic bacteria, including Liste-
ria and Shigella, use actin polymerization to move
through the cytoplasm of their animal cell hosts at about
0.5ms1 (Fig. 37.12A). These bacteria hijack the
machinery normally used to move the leading edge of
motile cells to polymerize a comet tail of actin filaments
that pushes the bacterium forward. One end of the bac-
terium has a concentration of proteins that directly (Lis-
teria) or indirectly (Shigella) activate Arp2/3 complex
to polymerize a network of branched actin filaments (see
Fig. 33.12). Growth of this network at its trailing end
pushes the bacterial cell forward. The comet tail of cross-
linked actin filaments is stationary and depolymerizes
distally at the same rate at which it grows next to the
bacterium, so it remains a constant length.
Vaccinia viruses attached to the outer surface of
animal cells move by a related mechanism. They use
transmembrane proteins to activate the cytoplasmic
actin assembly system to drive their movements at one
stage in its life cycle (Fig. 37.12B). Placement of a plastic
bead coated with adhesion proteins on the plasma mem-
brane of some animal cells can induce similar propulsive
actin comet tails in the cytoplasm.
Fungal and animal cells use Arp2/3 complex to assem-
ble actin filaments at sites of clathrin-mediated endocy-
tosis. Polymerization of these filaments assists with
vesicle separation from the plasma membrane. Nucle-
ation promoting factors associated with some endo-
somes stimulate Arp2/3 complex to assemble actin
filament comets and move similar to Listeria.
650 SECTION IX n Cytoskeleton and Cellular Motility

A. Listeria B. Vaccinia virus

5 m

FIGURE 37.12 Fluorescence micrographs of actin filament comet tails in animal epithelial cells infected with the bacterium Listeria monocy-
togenes (A) or vaccinia virus (B). Both pathogens are stained green with fluorescent antibodies. They use host cell proteins to assemble a
crosslinked network of actin filaments shaped like a comet tail. Actin filaments are stained red with rhodamine-phalloidin. A, The comet tail pushes
Listeria in a PtK cell through the cytoplasm and into projections of the plasma membrane at the edge of the cell. B, When the replicated vaccinia
viruses in this HeLa cell reach the cell surface 8 hours after infection, they activate Arp2/3 complex to assemble a cytoplasmic comet tail of actin
filaments that are thought to enhance the spread of the virus from cell to cell. Actin-based motility of vaccinia virus depends on tyrosine phos-
phorylation of a viral transmembrane protein A36R that remains inserted in the plasma membrane. (A, Courtesy K. Skoble, D. Portnoy, and M.
Welch, University of California, Berkeley. B, Courtesy T.P. Newsome and M. Way, Cancer Research UK, London. For reference, see Frischknecht
F, Moreau V, Rottger S, etal. Actin-based motility of vaccinia virus mimics receptor tyrosine kinase signaling. Nature. 1999;401:926929.)

ACKNOWLEDGMENTS Fu MM, Holzbaur EL. Integrated regulation of motor-driven organelle

We thank Anthony Brown, Erika Holzbaur, and Margaret
Titus for suggestions on revising this chapter.
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