CHAPTER 38

Cellular Motility

C ells move at rates that range over four orders of tubular pseudopods. Most cells, including white blood
magnitude (Fig. 38.1 and Table 38.1). At one extreme, cells, nerve growth cones, and fibroblasts move at
ciliates, bacteria, and sperm swim rapidly through water, intermediate rates.
and giant amoebas crawl rapidly over solid substrates. At Cells produce forces for motility in many different
the other extreme, fungal, algal, and plant cells with ways, most commonly using the same four mechanisms
rigid cell walls are immobile. However, even some that produce intracellular movements (see Chapter 37):
plant cells move, such as pollen, which extends contraction of actinmyosin networks, movement of
motors on microtubules, reversible assembly of actin fila-
ments, or reversible assembly of microtubules. These
mechanisms often complement each other, even where
Velocity (m/sec)
10-2 10-1 100 101 102 103
movement depends mainly on one system. For example,
E. coli microtubules contribute to actin-based pseudopod
extension by helping to specify the polarity of the cell.
Sperm This chapter compares these common mechanisms with
a few more unusual, but informative mechanisms: con-
Tetrahymena
traction of calcium-sensitive fibers of ciliates, reversible
assembly of novel cytoskeletal polymers of nematode
sperm, and rotation of bacterial flagellar motors.
Amoeba proteus Most cells possess all proteins required for cellular
motility, so the striking variation in their rates of move-
ment arises from differences in the abundance, organiza-
tion, and activities of this machinery. For example, both
White blood cell
nonmotile yeasts and contractile muscle cells contain
actin, myosin-II, heterodimeric capping protein, -actinin,
Pollen tube and tropomyosin. Yeasts use these proteins for cytokine-
sis (see Fig. 44.25), while muscle assembles high con-
Fibroblast centrations of similar proteins into sarcomeres (see
Figs. 39.2 and 39.3) for powerful, fast contractions.

Nerve Cell Shape Changes Produced by

FIGURE 38.1 VELOCITIES OF MOVING CELLS SPAN MORE
THAN FOUR ORDERS OF MAGNITUDE. Scale drawings of cells
with a range of velocities. E. coli, Escherichia coli.
Extension of Surface Processes
Alteration of cellular shape can be most simply brought
about by assembly of new cytoskeletal polymers or by
rearrangement of preexisting assemblies of actin fila-
ments or microtubules.

651
652 SECTION IX n Cytoskeleton and Cellular Motility

TABLE 38.1 Velocities of Cellular Movements

Unitary Velocity Summed Velocity
System (ms1) (ms1) Motile Mechanism
Striated muscle contraction (biceps) 510 48 105 Actin-myosin adenosine triphosphatase
Filopodium extension, Thyone sperm 10 10 Actin polymerization
Pseudopod extension, fibroblast 0.02 0.02 Actin polymerization
Pseudopod extension, human neutrophil 0.1 0.1 Actin polymerization
Pseudopod extension, Amoeba proteus ? 10 Actin-myosin ATPase
Pseudopod extension, nematode sperm 1 1 Assembly of major sperm protein
Retraction of axopodium, heliozoan >100 >100 Disassembly of microtubules
Spasmoneme contraction, Vorticella ? 23,000 Calcium-induced conformational change
Swimming, Escherichia coli 25 Flagellum powered by rotary motor
Swimming, sea urchin sperm 15 Microtubule-dynein ATPase

Note: Unitary velocity refers to a single molecular unit. Summed velocity is the overall motion of the cell.

EGG Filopodia
A B

JELLY
Egg stimulates Actin polymerization
secretion of extends the Bundle Ruffles
acrosomal contents acrosomal of actin
process filaments

Packet
of actin
and profilin
Nucleus
Axoneme 10 m

FIGURE 38.2 THYONE SPERM ACROSOMAL PROCESS. Actin
polymerization drives the growth of the acrosomal process of the
sperm of the sea slug, Thyone. The acrosome (red) is a membrane-
bound secretory vesicle, which fuses with the plasma membrane and
releases its hydrolytic enzymes prior to growth of the acrosomal
process. When the acrosomal process reaches the egg, the plasma
membranes of the two cells fuse. (Based on the work of L. Tilney,
University of Pennsylvania, Philadelphia.)
Studies of echinoderm sperm revealed that actin
polymerization drives the formation of cell surface pro-
jections called filopodia. To fertilize the egg the sperm
extends a long filopodium to penetrate the protective
jelly surrounding the egg (Fig. 38.2). Actin subunits for
this acrosomal process are stored with profilin (see Figs.
1.4 and 33.11) in a novel concentrated packet near the
nucleus. Contact with an egg stimulates actin filaments
to polymerize, starting from a dense structure near
the nucleus. Addition of subunits to the distal (barbed)
end of growing filaments drives the elongation of the
process and the surrounding membrane at a rate of 5 to
10ms1 (an astounding maximum of 3700 subunits per
second!). Actin subunits diffuse rapidly enough from
their storage site to drive this rapid elongation. The
number of filaments declines from approximately 150
FIGURE 38.3 FILOPODIA. A, Fluorescence micrograph of the
edge of a mouse NIH 3T3 cell expressing formin mDia2 and activated
Rif, a Rho-family guanosine triphosphatase (GTPase) that activates
mDia2. mDia2 concentrated at the tips of filopodia is stained red with
fluorescent antibodies. B, Scanning electron micrograph of mouse
macrophages spreading on a glass slide, illustrating the flat peripheral
lamellae, wave-like ruffles on the upper surface, and finger-like filo-
podia. (A, From Pellegrin S, Mellor H. The Rho family GTPase Rif
induces filopodia through mDia2. Curr Biol. 2005;15:129133.
B, From Chitu V, Pixley FJ, Macaluso F, etal. The PCH family member
MAYP/PSTPIP2 directly regulates F-actin bundling and enhances
filopodia formation and motility in macrophages. Mol Biol Cell. 2005;16:
29472959.)
near the base to less than 20 at the tip of the process,
presumably from capping. The molecules (if any) guiding
the growth of the filaments are not known.
Bundles of actin filaments support filopodia of a more
modest size on many other cells. Filopodia on macro-
phages (Fig. 38.3), nerve growth cones (Fig. 38.6A),
fibroblasts, and epithelial cells grow much more slowly
and depend on formins and vasodilator-stimulated
protein (VASP) at their tips (Fig. 38.3) to guide barbed-
end assembly of actin filaments crosslinked by a protein
called fascin. Microvilli of the brush border of epithelial
cells (see Fig. 33.2) are short, stable filopodia. The
 CHAPTER 38 n Cellular Motility 653

A B C

FIGURE 38.4 DYNAMIC CELL SURFACE PROJECTIONS SUPPORTED BY MICROTUBULES. AB, Drawings of the radiolarian Echi-
nospherium (a protozoan) showing projections called axopodia, which capture prey and draw them toward the cell body by transport on surface
of the projection or collapse of the projection. C, Electron micrograph of a thin section across an axopodium, showing the double spiral array of
microtubules. (Courtesy L. Tilney, University of Pennsylvania, Philadelphia.)

A. Epithelial folding Apical contraction B. Neural tube formation

folds the epithelium
Apex

Basal lamina Base

C. Drosophila ectoderm contraction

FIGURE 38.5 ACTOMYOSIN CONTRACTIONS MOLD THE SHAPE OF EPITHELIA DURING EMBRYONIC DEVELOPMENT. A, Folding
of a planar epithelium into a tube. B, Formation of the neural tube by contraction of the apical pole of columnar epithelial cells resulting in shape
change and invagination of the epithelium. C, Contraction around the margin of the ectoderm pulls this epithelium over the surface of a Drosophila
embryo. The scanning electron micrographs (SEMs [left and right]) show the steps in the dorsal closure of the epithelium. The time series of fluo-
rescence micrographs (center) shows live embryos expressing an actin-binding fragment of the protein moesin, fused to green fluorescent protein.
(C, SEMs courtesy Thom Kaufman, Indiana University, Bloomington [see his movie Fly Morph-o-genesis at http://www.sdbonline.org/archive/
dbcinema/kaufman/kaufman.html]; light micrographs courtesy D. Kiehart, Duke University, Durham, NC. For reference, see Kiehart DP, Galbraith
CG, Edwards KA, etal. Multiple forces contribute to cell sheet morphogenesis for dorsal closure in Drosophila. J Cell Biol. 2000;149:
471490.)

bundles of actin filaments supporting microvilli are
crosslinked to each other by fimbrin and villin and to
the plasma membrane by myosin-I. Synthesis of these
accessory proteins triggers assembly of microvilli on
epithelial cells as well as cells that normally have few
microvilli.
Some embryonic cells grow long filopodia to contact
cells micrometers distant. These filopodia create physical
contacts that allow the cells to send or receive signals
that influence developmental decisions. Long filopodia
between some pairs of cells even make gap junctions
(see Fig. 31.6).
654 SECTION IX n Cytoskeleton and Cellular Motility

A. Growth cone Lamellum Filopodia

n
Axo
B Leading Leading
edge edge

Keratocyte Cytoplast Continuous

gliding

C Protrusion
& adhesion

Tail
retraction Repeated cycles

FIGURE 38.6 MOTILITY BY PSEUDOPOD EXTENSION. A, Phase-contrast micrographs of a cultured nerve cells growth cone at 1-minute
intervals. The growth cone extends filopodia and fills in the space between with an actin-filled lamella. B, Gliding movements of a fish epidermal
keratocyte and a keratocyte cytoplast, a cell fragment consisting of the leading edge with most of the cell body including the nucleus removed.
Differential interference contrast micrographs at 15-second intervals were superimposed. Drawing shows the pattern of movement. C, Phase-
contrast micrograph of a keratocyte on glass. This cell moved toward the upper right using cycles of expansion of the broad leading lamella and
retraction of the trailing edge from the surface as shown by the drawings. (A, Courtesy D. Bray, University of Cambridge, United Kingdom.
B, From Pollard TD, Borisy GG. Cellular motility driven by assembly and disassembly of actin filaments. Cell. 2003;112:453465. C, Courtesy
J. Lee, University of Connecticut, Storrs.)

A group of protists called heliozoans, named for their
similarity to a cartoon of the sun, are a rare example of
using microtubules instead of actin filaments to extend,
support, and retract long, thin processes bounded by
the plasma membrane (Fig. 38.4). Microtubules in these
axopodia are crosslinked into a precise geometrical
array accounting for the rigidity of these long processes.
Axopodia capture prey organisms and transport them
toward the cell body for phagocytosis by two mecha-
nisms. Some move along the surface of axopodia (similar
to intraflagellar transport; Fig. 38.18). Other axopodia
collapse owing to rapid depolymerization of the micro-
tubules and drag the prey with them. Ca2+ influx appears
to trigger depolymerization of the microtubules, but the
details of the mechanism are not known.
Cell Shape Changes Produced
by Contraction
Cells can change shape by localized or oriented cyto-
plasmic contractions. Muscle contraction (Chapter 39)
and cytokinesis (Chapter 44) are the best examples, but
contractions also remodel many embryonic tissues. Local-
ized contractions at the base or apex of cells in a planar
epithelium cause evaginations or invaginations (Fig. 38.5)
such as those that form the neural tube (see Fig. 30.8)
and glands that bud from the gastrointestinal tract and
branches of the respiratory tract. Closure of the epider-
mis over a Drosophila embryo also requires contraction
of a circumferential ring of cells (Fig. 38.5C). Tension
generated by myosin-II and actin filaments deforms each
cell and, collectively, the whole epithelium. Similarly,
contraction of a ring of actin filaments associated with
the zonula adherens of intestinal epithelial cells helps
regulate the permeability of the tight junctions that seal
sheets of epithelial cells (see Fig. 31.2).
Locomotion by Pseudopod Extension
The ability to crawl over solid substrates or through
extracellular matrix is essential for many cells. Perhaps
the most spectacular example is the slowly moving
growth cone of a nerve axon (Fig. 38.6A). Although
moving less than 50nms1, growth cones navigate
 CHAPTER 38 n Cellular Motility 655

E. Growing filaments push membrane forward

Extracellular stimuli

G.
F. Capping protein

AT
C. WASp/Scars and existing

n
terminates

tio
P-h
70
filaments activate elongation

ga
Arp2/3

yd
Arp2/3 complex to

on
complex

rol
form a branch

El
ys

D.
B. Signals activate

is
WASp/Scar

an
proteins PAK

dP
i
dis
J. LIM-kinase

so
inhibits

cia
H. ADF/cofilin severs ADF / cofilin

tio
A. Pool of ATP-actin
ADP-actin filaments

n
bound to profilin
that depolymerize
ADF / cofilin
Profilin

Profilin/Actin
complexes
I. Profilin catalyzes exchange
of ADP for ATP

FIGURE 38.7 A MODEL FOR ACTIN FILAMENT ASSEMBLY AND DISASSEMBLY AT THE LEADING EDGE. The reactions are separated
in space for clarity but actually occur together along the leading edge. A, Cells contain a large pool of unpolymerized actin bound to profilin.
B, Stimulation of cell surface receptors produces activated Rho-family guanosine triphosphatases (GTPases) and other signals that activate
Wiskott-Aldrich syndrome protein (WASp)/Scar proteins. C, These proteins, in turn, activate nucleation of new actin filaments by Arp2/3 complex
on the side of existing filaments. D, The new filaments grow at their barbed ends until they are capped (see F). E, Growing filaments push the
plasma membrane forward. F, Capping protein terminates elongation. G, Polymerized adenosine triphosphate (ATP)-actin (yellow) hydrolyzes the
bound adenosine triphosphate (ATP) to adenosine diphosphate (ADP) and inorganic phosphate (Pi) (orange), followed by slow dissociation of
phosphate yielding ADP-actin (red). H, ADF/cofilins bind and sever ADP-actin filaments and that disassemble, releasing ADP-actin. I, Profilin
promotes the exchange of ADP for ATP, restoring the pool of unpolymerized ATP-actin bound to profilin. J, Some of the same stimuli that initiate
polymerization can also stabilize filaments when LIM-kinase phosphorylates actin-depolymerizing factor (ADF)/cofilins, inhibiting their depolymer-
izing activity. Inset, Electron micrograph of the branched network of actin filaments at the leading edge. PAK, p21-activated kinase. (Modified
from Pollard TD, Blanchoin L, Mullins RD. Biophysics of actin filament dynamics in nonmuscle cells. Annu Rev Biophys Biomol Struct.
2000;29:545576. Inset, Courtesy T. Svitkina and G. Borisy, Northwestern University, Evanston, IL.)

precisely over distances ranging from micrometers to
meters to establish all the connections in the human
nervous system, which consists of billions of neurons
and approximately 1.6 million kilometers of cellular
processes. Some epithelial cells (Fig. 38.6B) and white
blood cells move much faster, about 0.5ms1. These
movements enable epithelial cells to cover wounds and
allow leukocytes to move from the blood circulation to
sites of inflammation (see Fig. 30.13) where they engulf
microorganisms by phagocytosis (see Fig. 22.3). During
vertebrate embryogenesis, neural crest cells migrate long
distances through connective tissues before differentiat-
ing into pigment cells and sympathetic neurons. Fibro-
blasts lay down collagen fibrils as they move through the
extracellular matrix (see Fig. 29.4).
The best-characterized motile system is animal and
amoeboid cells moving on a flat surface such as a micro-
scope slide that may be coated with extracellular matrix
molecules. These cells use actin polymerization to
extend the leading edge, then adhere to the underlying
substrate, and (if the whole cell is to move) release and
retract any attachments of its tail to the substrate. Animal
cells move in other environments, such as three-
dimensional extracellular matrix, using variations of this
theme as described at the end of the section.
Lamellar Motility on Flat Surfaces by
Pseudopod Extension
Pseudopods that lead the way in cell migration are filled
with a dense network of branched actin filaments with
their fast-growing barbed ends generally facing out
towards the plasma membrane (Fig. 38.7). The leading
lamella is autonomous for locomotion, and moves nor-
mally after amputation from the rest of the cell (Fig.
38.6B). Generally, the leading lamella is very flat, on the
order of 0.25m thick, but some cells extend the lamellae
up from the substrate into a wave-like fold called a ruffle
(Fig. 38.3). Microtubules help maintain the polarized
656 SECTION IX n Cytoskeleton and Cellular Motility
A 0s 24 s 77 s 133 s 274 s
5 m
B 4s 48 s 81 s
10 m
FIGURE 38.8 DOCUMENTATION OF ACTIN FILAMENT
DYNAMICS AT THE LEADING EDGE WITH FLUORESCENT
ACTINS. A, Fluorescence photobleaching experiment with a stationary
cell. Fluorescent actin is injected into a cultured epithelial cell and
allowed to incorporate into filaments. A laser pulse bleaches some of
the fluorescent actin, leaving a dark spot (arrow) that reveals movement
of the filaments toward the cell center. The spot recovers fluorescence
as diffusing fluorescent actin assembles new filaments in the bleached
zone. B, Caged fluorescent actin experiment with a motile cell. Fluo-
rescent dye bound to actin is masked with a chemical group preventing
fluorescence. After incorporation into actin filaments of a fish keratocyte
(see Fig. 33.2E), dyes in one area of the cell are uncaged with a light
pulse (arrow), and red fluorescence is followed with time. Fluorescent
actin filaments are stationary with respect to the substrate as the cell
moves forward (upward). The fluorescent spot of marked filaments
fades with time, owing to depolymerization and dispersal of the fluo-
rescent subunits. (A, From Wang Y-L. Exchange of actin subunits at
the leading edge of living fibroblasts: possible role of treadmilling.
J Cell Biol. 1985;101:597602, copyright The Rockefeller University
Press. B, From Theriot JA, Mitchison TJ. Actin microfilament dynamics
in locomoting cells. Nature. 1991;352:126131.)
shape that is required for persistent directional locomo-
tion, but are not required for pseudopod extension.
Actin filaments assemble continuously near the
leading edge of pseudopods and turn over rapidly
deeper in the cytoplasm (Fig. 38.8). Thus, the inhibitor
cytochalasin stops actin polymerization at the leading
edge (see Fig. 33.18). Purified actin was labeled with a
fluorescent dye and microinjected into live cells, where
it incorporated into actin-containing structures, includ-
ing the cortical network, pseudopods, stress fibers, and
surface microspikes. Photobleaching (Fig. 38.8A) or
photoactivating the fluorescent actin (Fig. 38.8B) showed
that actin assembles at the leading edge and then turns
over. Alternatively, if a low concentration of fluorescent
actin is injected it will incorporate irregularly into fila-
ments, producing tiny speckles of fluorescence that
can be tracked over time to reveal sites of assembly and
disassembly (see Fig. 33.18DE).
Arp2/3 complex (see Fig. 33.12) and formins
cooperate with VASP (see Fig. 33.14) to initiate and
elongate actin filaments at the leading edge of motile
cells. Chemotactic stimuli (Fig. 38.10) or intrinsic signals
transduced by Rho-family guanosine triphosphatases
(GTPases), membrane polyphosphoinositides, and
proteins with SH3 (Src homology 3) domains activate
Wiskott-Aldrich syndrome protein (WASp)/Scar
proteins (see Fig. 33.13), which promote the forma-
tion of actin filament branches by Arp2/3 complex. The
pool of unpolymerized actin maintained by profilin (and
thymosin-4, where it is present) drives the elongation of
actin filament branches at 50 to 500 subunits per second.
Growing filaments are generally oriented toward the
leading edge and push against the inside of the plasma
membrane with forces in the piconewton (pN) range.
These forces bend the filaments, which favors branching
on their convex surface, facing the leading edge. Het-
erodimeric capping protein (see Fig. 33.15) terminates
elongation of these branches before they grow longer
than 1m. Longer filaments are less effective at pushing,
since they buckle under piconewton forces. A similar
mechanism assembles actin filaments in comet tails on
intracellular bacteria and viruses (see Fig. 37.12) and
endocytic actin patches in yeast (see Figs. 6.3 and 37.11).
The recycling of actin and accessory proteins is
essential for thousands of rounds of assembly as the cell
moves forward. Actin-depolymerizing factor (ADF)/
cofilins (see Fig. 33.16) bind and sever aged adenosine
diphosphate (ADP)-actin filaments located away from
the leading edge. The fragments may be capped, in
which case they slowly depolymerize at the pointed
ends. This recycling mechanism is so efficient that the
branched network is disassembled in seconds.
Formins assemble long unbranched filaments (see Fig.
33.2E) in the flat region of the cell just behind the
branched network at the leading edge. Some of these
filaments form bundles that protrude as filopodia beyond
the leading edge (Fig. 38.3), where VASP (see Fig. 33.14)
promotes their elongation. The tropomyosin binds along
the sides of these long filaments, protecting them from
severing by ADF/cofilins.
Actin filament crosslinking proteins stabilize pseudo-
pods. Human melanoma cells that lack the crosslinking
protein filamin (see Fig. 33.17) form unstable pseudo-
pods all around their peripheries and locomote abnor-
mally (Fig. 38.9). These tumor cells recover their normal
behavior if provided with filamin. Similarly, Dictyoste-
lium cells that lack a homolog of filamin form fewer
pseudopods.
Influence of the Substrate on Lamellar Motility
The growing actin network at the leading edge will either
push the membrane forward or slip backward depending
on how well it is connected to the substrate across the
plasma membrane. In highly motile cells such as epithelial
cells from fish scales (Fig. 38.6B) transmembrane
adhesion proteins anchor the actin filament network
to the substrate, so the polymerization results in forward
motion (Fig. 38.8B). In stationary cells (Fig. 38.8A;

A B
Migration
FIGURE 38.9 CONTRIBUTION OF THE ACTIN FILAMENT
CROSSLINKING PROTEIN FILAMIN TO THE STABILITY OF THE
LEADING EDGE OF HUMAN MELANOMA CELLS. Pairs of phase-
contrast light micrographs, taken at different times, of living cells grown
in serum-containing medium on a plastic surface. A, Melanoma cells
expressing filamin have normal leading lamella. B, Melanoma cells
lacking filamin form spherical blebs around their margins and migrate
very little. (Courtesy C. Cunningham and T.P. Stossel, Harvard Medical
School, Boston, MA. For reference, see Cunningham C, Gorlin JB,
Kwiatkowski DJ, etal. Actin-binding protein requirement for cortical
stability and efficient locomotion. Science. 1992;255:325327.)
also see Fig. 33.18DE), actin polymerizes at the edge of
the cell causing the entire network to move en masse
away from the membrane, a phenomenon called retro-
grade flow. Fibroblasts are an intermediate state, in which
actin polymerization produces some forward movement
but also considerable retrograde flow.
Movement driven by adhesion requires a compromise.
Adhesion must be strong enough for the internal forces
to propel the cell forward but not so strong that it pre-
vents movement. Transmembrane adhesion proteins
such as integrins (see Fig. 30.9) both anchor the cell and
transmit the presence of their ligands and the stiffness
of the environment to Rho-family GTPases that regulate
actin assembly and myosin contractility. Rapidly revers-
ible binding of integrins and other adhesion proteins to
extracellular matrix molecules, such as fibronectin,
allows adhesion without immobilization. Rapidly moving
white blood cells attach weakly and transiently, whereas
slowly moving fibroblasts form longer-lasting focal con-
tacts (see Fig. 30.11). Cells tend to move up gradients of
adhesiveness but stop if adhesion is too strong. This
graded response to adhesion allows neural crest cells to
migrate preferentially through regions of embryonic
connective tissue marked by adhesive proteins.
Tail Retraction and Other Roles for Myosin
in Motility
Growth cones of neurons draw out a long process from
a stationary cell body, but most cells must break adhe-
sions at their trailing edge to advance. Adherent, slowly
moving cells such as fibroblasts exert significant tension
CHAPTER 38 n Cellular Motility 657
on the underlying substrate, when myosin pulls on the
bundles of actin filaments associated with focal contacts
(see Fig. 30.11). When tension overcomes the attach-
ments, the rear of the cell shortens elastically and then
contracts further (Fig. 38.6C). Myosin also contributes to
the retrograde flow of actin filaments in the zone between
the leading edge and the cell body.
Other Modes of Motility
Cells that move in two-dimensions by pseudopod exten-
sion and tail retraction repurpose the same proteins to
behave differently in other environments. In the extra-
cellular matrix of connective tissues (see Chapter 29)
cells must squeeze between three-dimensional networks
of fibers using forces produced by myosin to generate
hydrostatic pressure to push forward and to pull the
relatively stiff nucleus forward. In other circumstances
cells can expand by forming blebs of plasma membrane
lacking networks of actin filaments. Nerve growth cones
(Fig. 38.6A) extend filopodia and then fill the spaces in
between them with a lamella containing new, branched
actin filaments. Superfast giant amoebas (Fig. 38.1) use
myosin to generate contractions in the cortex or the
front of the pseudopod to drive the bulk streaming of
cytoplasm into advancing pseudopods.
Chemotaxis of Motile Cells
Extracellular chemical clues direct locomotion by influ-
encing the formation and persistence of pseudopods.
Movement toward a positive signal is called chemotaxis.
The best-characterized example is the attraction of Dic-
tyostelium to cyclic adenosine monophosphate (cAMP)
(Fig. 38.10), the extracellular chemical that these
amoebas use to communicate as they form colonies
before making spores. Remarkably, these cells can sense
a gradient of cAMP corresponding to a concentration
difference of less than 2% along their length. This small
difference is amplified into strong internal signals that
control motility. Binding of cAMP to seven-helix recep-
tors in the plasma membrane activates trimeric G-proteins
inside the cell (see Fig. 25.9). The G-proteins activate
pathways that regulate the activity of enzymes that
control the concentration of the lipid second messenger
phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the
plasma membrane: phosphatidylinositol 3-kinase (PI3K)
synthesizes PIP3 and PTEN (phosphatase and tensin
homolog) degrades it. (See Fig. 26.7 for details on poly-
phosphoinositides.) A fast positive pathway that is sensi-
tive to local receptor occupancy and a slower global
negative signal that is proportional to total receptor
occupancy produce a gradient of PI3K activity inside the
cell that is highest near the external source of cAMP.
These pathways concentrate PTEN on the plasma mem-
brane away from the source of cAMP. This complemen-
tary regulation of the kinase and phosphatase creates an
internal gradient of PIP3 three to seven times steeper
658 SECTION IX n Cytoskeleton and Cellular Motility

A. Time course of turning toward gradient of cAMP

0 15 30 45 60 75 90 105 120 135 150

B. Amplification of the shallow external gradient of cAMP C. Concentration of plasma

into a sharp biochemical gradient inside the cell: membrane PIP3 toward
Source the source of cAMP
of cAMP
Shallow gradient
of cAMP outside Membrane
Receptor

Gradient of
PIP3 on inside
of plasma
membrane

0 sec 8 sec 20 sec

Fast, local excitation Slow, global inhibition Response

FIGURE 38.10 CHEMOTAXIS OF A DICTYOSTELIUM AMOEBA TOWARD CYCLIC ADENOSINE MONOPHOSPHATE. A, Live cell
attracted to cyclic adenosine monophosphate (cAMP) (gold) released from a micropipette. A time series of differential interference micrographs
shows the rapid formation of a new pseudopod and reorientation of the direction of movement when the position of the micropipette is moved
at the 60-second time point. B, Cells have a uniform distribution of cAMP receptors (yellow and red dots) over their surface. A shallow gradient
of cAMP activates these seven-helix receptors (red), which activate a trimeric G-protein and phosphatidylinositol 3-kinase, an enzyme that rapidly
converts phosphatidylinositol 4-phosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3). On a slower time scale, the active G-protein
activates phosphatase and tensin homolog (PTEN), a PIP3 phosphatase, throughout the cell. The combination of these two signals creates a
steep gradient of PIP3 across the cell. C, Fluorescence micrograph of a cell exposed to a point source of cAMP (yellow). A green fluorescence
protein (GFP)-pleckstrin homology (PH) domain fusion protein (green) inside the cell binds to PIP3 on the inside of the plasma membrane, revealing
the steep gradient of PIP3. (A, Courtesy Susan Lee and Richard Firtel, University of California, San Diego. B, Modified from a sketch by Pablo
Iglesias, Johns Hopkins University, Baltimore, MD. C, Courtesy Pablo Iglesias, Johns Hopkins University, Baltimore, MD. For reference, see
Janetopoulos C, Ma L, Devreotes PN, Iglesias PA. Chemoattractant-induced phosphatidylinositol 3,4,5 trisphosphate accumulation is spatially
amplified and adapts, independent of the actin cytoskeleton. Proc Natl Acad Sci U S A. 2004;101:89518956.)

than the external gradient of cAMP. Transduction of this
internal gradient of PIP3 into motility requires Rho-family
GTPases and formation of new actin filaments. Local
polymerization and crosslinking of these actin filaments
expand the cortex facing the source of cAMP into a new
pseudopod and move the cell toward the cAMP.
Leukocytes are attracted to chemokines and bacterial
metabolites at sites of infection (see Fig. 30.13), espe-
cially small peptides derived from the N-termini of
bacterial proteins, such as N-formyl-methionine-
leucine-phenylalanine (referred to as FMLP in the
scientific literature). Similar to Dictyostelium, activation
of seven-helix receptors and trimeric G-proteins ampli-
fies shallow external gradients of FMLP into steeper
internal gradients of PIP3 and other signals that control
pseudopod formation.
Negative signals also influence pseudopod persistence
and the direction of motility. A classic example is the
negative effect of contact with another cell. Loss of
contact inhibition of motility by tumor cells contrib-
utes to their tendency to migrate among other cells and
spread throughout the body.
Growth Cone Guidance: A Model for Regulation
of Motility
Growth cones of embryonic nerve cells use a combina-
tion of positive and negative cues to navigate with high
reliability to precisely the right location to create a
synapse (Fig. 38.11). This combinatorial strategy is much
more complex than the simple chemoattraction of Dic-
tyostelium to cAMP, as expected for the more compli-
cated task of connecting billions of neurons to each
other or specific muscle cells. Cues for growth cone
guidance come from soluble factors and cell surface
molecules, each with a specific receptor on the growth
cone. As in other systems, extracellular matrix molecules
provide the substrate for growth cone movements. Pre-
cisely positioned expression of cue molecules and their
 CHAPTER 38 n Cellular Motility 659

A
receptors, such as members of the DCC family, steer the
growth cone toward a netrin source by activating actin
polymerization by VASP. Other netrin receptors repel
the growth cone from netrin. Growth cones without
these receptors are insensitive to this cue.
Two commissures Slit, a large extracellular matrix protein, repels growth
per segment
cones with Slit receptors, which are immunoglobulin
cell adhesion molecules (IgCAMs) called Robo1, Robo2,
and Robo3. Mutations in the genes for these receptors
cause growth cones to ignore Slit.
Longitudinal IgCAM cell surface adhesion proteins (see Fig. 30.3),
axon bundles such as fasciculin II, prompt growing axons to bundle
together in bundles called fascicles by homophilic inter-
actions. Growth cones can be attracted out of these
bundles to particular targets, such as muscle cells, that
Midline secrete chemoattractants or proteins that antagonize
fasciculin II adhesion.
Netrin
The effects of mutations in genes for receptors and
B
gradient their ligands revealed how growth cones in Drosophila
embryos navigate using multiple guidance cues (Fig.
Neurons with Slit in matrix repels 38.11). Growth cones of neurons on one side of the
ipsilateral growth cones nerve cord migrate across the midline to the opposite
projections with Robo1
side and then navigate faithfully to their targets. Netrins
High Robo1 repels secreted by cells at the midline attract growth cones
growth cone from
midline expressing the DCC (Frazzled) netrin receptor. However,
midline cells also secrete high levels of the matrix protein
Frazzled receptor
(DCC) for netrin attracts Slit, which repels growth cones. Growth cones cross the
growth cone to midline midline by downregulating the slit receptor. Once across
the midline, the growth cones upregulate the slit recep-
Comm on glial cells
downregulates tor, so they never cross back to the side of origin. Local
Robo1 expression and cues alert particular growth cones of motor neurons to
allows growth cone
to pass midline branch off of fascicles to innervate individual muscle
cells. Path finding by capillaries uses some of the same
High Robo1 drives
growth cone out of guidance mechanisms to grow blood vessels.
midline and prevents
recrossing
Eukaryotic Cilia and Flagella
FIGURE 38.11 DROSOPHILA GROWTH CONE GUIDANCE.
A, Light micrograph of a filleted embryo showing the nerve cord Axonemes built from microtubules and powered
stained brown with an axon marker. The axons of approximately 90% by dynein produce the beating of cilia and flagella
of neurons cross the midline a single time in a transverse nerve bundle (Fig. 38.12). These exceedingly complex structures are
called a commissure before running longitudinally in fascicles on each remarkably ancient. More than 1 billion years ago the last
side of the midline. B, Drawing showing the ligands and receptors that
common eukaryotic ancestor (see Fig. 2.4) had motile
guide growth cones across the midline and prevent their return to the
ipsilateral (original) side. Frazzled receptors for netrin attract the growth flagella with all the essential features of human cilia and
cone to the midline where Comm downregulates the activity of Robo1, flagella, as evidenced by motile axonemes in most
a repulsive receptor for Slit, allowing axons to cross the midline. branches of eukaryotes. However, most fungi and plants
(A, Courtesy John Thomas, Salk Institute, La Jolla, CA.) lost the genes for axonemal proteins.
Animal cells make three types of axonemes, all with
nine outer doublet microtubules anchored by a basal
receptors guides growth cones along a staggering body at the cortex of the cell and surrounded by
number of different pathways, as illustrated by the fol- plasma membrane (Fig. 38.12B). Motile axonemes have
lowing well-characterized examples. dynein motors, a central pair of single microtubules
Localized cells in the nervous system, such as those and radial spokes (Figs. 38.14 and 38.15). Rotating nodal
in the floor plate of the developing spinal cord, secrete cilia (see below for the biological context) have dynein
soluble guidance proteins such as netrin. Gradients arms but no central pair or radial spokes. Immobile
of netrin provide long-range guidance for growth cones primary cilia lack dynein, central pairs and radial spokes.
of cells that possess netrin receptors. Some netrin This section begins with motile cilia and flagella.
660 SECTION IX n Cytoskeleton and Cellular Motility

A. Flagella B. Cilia Ciliary motion Fluid motion No motion

Cell motion
Effective
Flagellar motion stroke

Recovery
stroke

Cell motion Beating Rotary Primary

C. Cilia metachronal wave

9 + 2 + dynein + radial spokes 9 + 0 + dynein 9+0

Adapted from P. Satir

FIGURE 38.12 BEATING PATTERNS OF CILIA AND FLAGELLA. A, Waves of a sperm flagellum. B, Behavior of three types of cilia with
cross sections of their axonemes below. Left, Ciliary power and recovery strokes of a beating cilium. Middle, Rotary cilium. Right, Primary cilium.
C, Coordinated beating of cilia on the surface of an epithelium. (Modified from a drawing by P. Satir, Albert Einstein College of Medicine,
New York, NY.)

Properties of Cilia and Flagella A

10 m
Cilia and flagella are distinguished from each other by
their beating patterns (Fig. 38.12), but are nearly identi-
cal in structure. In fact, the flagella of the green alga
Chlamydomonas can alternate between propagating
waves typical of flagella and the oar-like rowing motion
of cilia. So one can use the terms cilia and flagella inter-
changeably. Subtle differences in the mechanism that
converts the dynein-powered sliding of the axonemal B
microtubules into movements determine which beating
pattern is produced.
Both cilia and flagella can propel cells as they cycle
rapidly, beating up to 100 times per second. Propagation
of bends along the length of individual flagella pushes a cell
such as sperm forward (Figs. 38.1 and 38.15). Coordinated
beating of many cilia can also move large cells (Fig. 38.13A).
Reversal of the direction of the power stroke allows a
unicellular ciliate to swim forward or backward. Alterna- FIGURE 38.13 IMAGES OF CILIA. A, Scanning electron micro-
graph of Paramecium showing waves of effective strokes passing
tively, if the cell is immobilized, like the epithelial cells regularly over the cell surface from one end to the other to keep the
lining an animal respiratory tract or forming the embryonic cell moving steadily forward. B, Fluorescence micrograph of cilia
skin (Fig. 38.13B), coordinated beating of cilia propels fluid (green, stained with fluorescent antibodies to tubulin) on epithelial cells
and particles over their apical surface. Ctenophores fuse of frog skin also stained with Alexa 647-phalloidin for actin filaments.
the membranes of many cilia together to make macrocilia (A, Courtesy T. Hamasaki, Albert Einstein College of Medicine, New
York, NY. From Lieberman SJ, Hamasaki T, Satir P. Ultrastructure and
that propel the organism in a manner similar to that of fins. motion analysis of permeabilized Paramecium. Cell Motil Cytoskeleton.
9:7384, 1988. B, Courtesy Brian J Mitchell of Northwestern Univer-
Structure of the Axoneme sity. From Werner ME, Hwang P, Huisman F, et al. Actin and microtu-
The nine outer doublet microtubules of all axonemes bules drive differential aspects of planar cell polarity in multiciliated
consist of one complete A-microtubule with the usual 13 cells. J Cell Biol. 2011;195:1926.)
protofilaments, bearing an incomplete B-microtubule
composed of 10 protofilaments attached to its side by but elastic. Pioneering genetic analyses and more recent
distinct junctional structures on either side. The filamen- proteomic studies established the locations of many of
tous protein tektin associates with one protofilament in these polypeptides, such as the 17 proteins that make
the wall of the A-microtubule. The central pair are up the radial spokes between the central sheath and
typical 13-protofilament microtubules. The plus ends of the outer doublets and the 11 protein subunits of the
all axonemal microtubules are at the distal tip. nexin-dynein regulatory complex that links outer dou-
More than 200 accessory proteins associate with the blets to each other. A long coiled-coil protein acts as a
9 + 2 microtubules (Fig. 38.14A), making axonemes stiff molecular ruler to specify the longitudinal positions of
 CHAPTER 38 n Cellular Motility 661

D E
A *
**
Large Bt B B-tubule
*
*
* At
ODA
** A A-tubule
*

* * *
() * (+)
N-RDC 24 nm
* IDA Spoke
*
* * S1
Small 24 nm
Outer
B C. Cilium with axoneme viewed from tip dynein
arm S2
32 nm
Outer doublet 96 nm
microtubule
S3
Membrane Inner 40 nm
Radial spoke dynein
arm
Central sheath S4
Central singlet tubule
Link to membrane
FIGURE 38.14 COMPOSITION AND STRUCTURE OF THE AXONEME. A, Two-dimensional gel electrophoresis separating more than 100
polypeptides of the axoneme of Chlamydomonas. Marked polypeptides (blue asterisks) are components of radial spokes. B, Electron micrograph
of a thin cross section of a ciliary axoneme stained with tannic acid. C, Drawing of a cross section of a cilium. D, Three-dimensional reconstruction
of one outer doublet and radial spoke based on electron microscopy tomography showing inner (IDA) and outer dynein arms (ODA) and a
nexin-dynein regulatory complex (N-DRC; blue). E, A short section of an outer doublet. In this example, the outer arm dyneins have two heads.
In some species, they have three heads. The dimensions indicate the longitudinal spacing between dynein arms and radial spokes. (A, Courtesy
B. Huang, Scripps Research Institute, La Jolla, CA. B, Courtesy R. Linck, University of Minnesota, Minneapolis. D, From Song K, Awata J,
Tritschler D, Bower R, et al. In situ localization of N and C termini of subunits of the flagellar nexin-dynein regulatory complex (N-DRC) using
SNAP tag and cryo-electron tomography. J Biol Chem. 2015;290:53415353. E, From a Chlamydomonas axoneme modified from Amos LA,
Amos WB. Molecules of the Cytoskeleton. New York: Guilford Press; 1991.)

many of these proteins with a 96nm periodicity along
the outer doublet microtubules. Central pair microtu-
bules are connected by a bridge and decorated by elabo-
rate projections. Mutations of these genes compromise
axonemal function in experimental animals and human
patients.
A family of axonemal dyneins bound to outer dou-
blets generates force for movement. Each dynein consists
of a large heavy chain forming a globular AAA ATPase
(adenosine triphosphatase) domain and a tail anchored
to an A-tubule by light and intermediate chains. A thin
stalk projecting from the catalytic domain exerts force
on the adjacent B-tubule during part of the ATPase cycle
(see Figs. 36.14 and 36.15). Like other axonemal proteins
the pattern of dynein arms repeats every 96nm along
the A-tubule of each outer doublet (Fig. 38.14D). The
outer row of dynein arms consists of four copies of
three-headed molecules in each repeat. The inner dynein
arms are more complicated: seven different dynein heavy
chains form six arms with one head plus one arm with
two heads in each repeat.
Mechanism of Axoneme Bending
Dynein-powered sliding of outer doublets relative to
each other bends axonemes. Sliding was first inferred
from electron micrographs of the distal tips of
microtubules in bent cilia. Later, sliding was observed
directly by loosening connections between outer dou-
blets with proteolytic enzymes and then adding adenos-
ine triphosphate (ATP) to allow dynein to push the
microtubules past each other (Fig. 38.15B). Sliding can
be followed in axonemes stripped of their membrane
by marking outer doublets with small gold beads (Fig.
38.15A). As outer doublets slide past each other, the
relative positions of the beads change. Dynein attached
to one doublet walks toward the base of the adjacent
microtubule, pushing its neighbor toward the tip of
the axoneme.
Biochemical extraction or genetic deletion of specific
dynein isoforms alters the frequency and waveform of
axonemal bending. Inner dynein arms are required for
flagellar beating, and deletion of even a single type of
inner-arm dynein can alter the waveform. Outer dynein
arms are not essential but influence the beat frequency
and add power to the inner arms. Humans with Karta-
gener syndrome lack visible dynein arms and have
immotile sperm and cilia. As a result, affected males are
infertile, and both men and women have serious respira-
tory infections, owing to poor clearance of bacteria and
other foreign matter from the lungs.
The mechanism of beating is intrinsic to the axoneme.
Thus, sperm tail axonemes swim normally when provided
662 SECTION IX n Cytoskeleton and Cellular Motility

A B

FIGURE 38.15 SLIDING MOVEMENTS OF OUTER DOUBLETS OF AXONEMES. AB, Time series of darkfield light micrographs. A, Sea
urchin sperm extracted with the detergent Triton X-100 and reactivated with ATP. Gold microbeads attached to two different outer doublets allow
the visualization of their displacement as the tail bends. B, Fragment of a sea urchin flagellar axoneme treated with trypsin. The addition of ATP
results in outer doublets sliding past each other out of the ends of the axonemal fragment. C, Electron micrograph of two outer doublets that
have slid past each other in an experiment similar to that in panel B. (A, From Brokaw CJ. Microtubule sliding in swimming sperm flagella. J Cell
Biol. 1991;114:12011215, copyright The Rockefeller University Press. B, Courtesy Ian Gibbons, University of California, Berkeley. For more
information, see Summers KE, Gibbons I. ATP-induced sliding of tubules in trypsin-treated flagella of sea-urchin sperm. Proc Natl Acad Sci
U S A. 1971;68:30923096. C, Courtesy P. Satir, Albert Einstein College of Medicine, New York, NY. For more information, see Sale WS,
Satir P. Direction of active sliding of microtubules in Tetrahymena cilia. Proc Natl Acad Sci U S A. 1977;74:20452049.)

with ATP, even without the plasma membrane or soluble
cytoplasmic components (Fig. 38.15A). Experiments
with these demembranated sperm models revealed
that the dynein ATPase activity is tightly coupled
to movement. The beat frequency is proportional to
ATPase activity, regardless of whether the frequency is
limited by increasing the viscosity of the medium or
the enzyme activity is limited by decreasing the ATP
concentration.
The bending that produces the bending waves of
flagella or the power and recovery strokes of cilia results
from local variation in the rate of sliding of pairs of outer
doublet microtubules along the length of an axoneme.
Coordination of these events is still being investigated,
but at least two factors are involved. Mutations show that
the central pair and radial spokes help coordinate the
activity of the dyneins around the circumference of
the axoneme as it bends. Mechanical constraints are
also required to convert microtubule sliding into local
bending. Destruction of the links between outer doublets
frees them to slide past each other rather than bending
the axoneme.
Although axonemes function autonomously, signal
transduction pathways regulate their activities. Photo-
taxis of Chlamydomonas is a particularly clear example
of how fluctuations in intracellular Ca2+ can modify fla-
gellar activity. The release of Ca2+ affects the two flagella
of the organism differentially and allows a cell to steer
toward or away from light (Fig. 38.16). Ciliates also have
mechanosensitive channels that depolarize the plasma
membrane when the organism collides with something.
Depolarization opens voltage-sensitive plasma membrane
Ca2+ channels, admitting Ca2+ into the cell. This reverses
the direction of ciliary beat. Both calcium and cAMP-
dependent phosphorylation of outer-arm dynein can
change the beat frequency (all the way to zero) or alter
the waveform.
Basal Bodies and Axoneme Formation
The mother centriole matures into a basal body that
anchors and templates the axoneme (Fig. 38.17; see also
Fig. 34.3B). After migrating to the cortex during inter-
phase, its distal appendages dock on the plasma mem-
brane and the nine outer doublet microtubules of the
axoneme grow directly from the nine outer triplet micro-
tubules of the basal body. This differs from microtubule
nucleation in the pericentriolar material during inter-
phase (see Fig. 34.15). Some protozoa use basal bodies
as centrioles during mitosis. Multiciliated cells form a
basal body for each axoneme de novo. Basal bodies are
typically anchored by protein fibers called rootlets. Cilia
seem to function normally in mice lacking the major
rootlet protein, but are unstable over the long term.
Mutations of genes for basal body proteins compromise
axonemal function in experimental animals and cause
human diseases.
 CHAPTER 38 n Cellular Motility 663

A B
A. Normal forward
swimming

B. Phototaxis D. Photoshock

C Central
h h singlet
micro-
Ca2+ tubule
Ca2+

Membrane

Outer
doublet
micro-
tubules

Matrix

Triplet
microtubules

C. Normal swimming parallel to new light direction

Cartwheel
stucture

FIGURE 38.16 CHLAMYDOMONAS PHOTOTAXIS. A, Normal
swimming toward the light using a cilia-like rowing motion of the fla-
gella. Absorption of light by a sensory rhodopsin (related to sensory
rhodopsins in Archaea) in the eyespot keeps the cell oriented.
B, Moderate-intensity light from the side causes Ca2+ to enter the
cytoplasm from outside the cell. The two flagella react differently,
causing the cell to turn toward the light. C, Once the cell is reoriented,
the flagella beat equally, and the cell swims toward the light. D, High-
intensity light releases a high concentration of Ca2+ and causes transient
wave-like motion of the flagella. This backward swimming allows the
cell to reorient and to swim away from the light.
FIGURE 38.17 BASAL BODIES. A, Electron micrograph of a
thin cross section of a basal body. B, Electron micrograph of thin
longitudinal section of basal bodies and proximal axonemes of cilia.
C, Drawings of cross sections and a three-dimensional (3D) drawing
of the basal body and proximal flagella of Chlamydomonas. In the 3D
drawing, the near side outer doublets are cut away to reveal the central
pair microtubules. (AB, Courtesy D.W. Fawcett, Harvard Medical
School, Boston, MA. C, Modified from Amos LA, Amos WB. Molecules
of the Cytoskeleton. New York: Guilford Press; 1991.)
664 SECTION IX n Cytoskeleton and Cellular Motility

A C D
Turnaround
Regenerating, zone
short flagella Axonemes
Tagged grow at
tubulin distal tips

Anterograde
IFT particle
Kinesin 2
Mate 2 hours

Dynein
Fusing cell Fused cell

IFT particle
Retrograde
Cut

Cell
body

FIGURE 38.18 FLAGELLAR GROWTH AND INTRAFLAGELLAR TRANSPORT. A, Incorporation of protein subunits at the tip of growing
Chlamydomonas flagella is revealed by an experiment involving the fusion of two cells, one expressing tubulin with an epitope tag that reacts
with a specific antibody and the other regenerating its flagella. As is shown in the fluorescence micrograph, tagged tubulin is incorporated only
at the distal tips of the growing flagella. Cells with paralyzed flagella made this experiment more convenient. B, Time course of regeneration of
Chlamydomonas flagellum following amputation of one flagellum. The surviving flagellum shortens transiently before both grow out together.
C, Electron micrographs of thin sections of Chlamydomonas flagella showing intraflagellar transport particles (arrows). D, Model for intraflagellar
transport (IFT). (A, Courtesy K. Johnson, Haverford College, Haverford, PA. Inset, From Johnson KA, Rosenbaum JL. Polarity of flagellar assembly
in Chlamydomonas. J Cell Biol. 1992;119:16051611, copyright The Rockefeller University Press. B, Based on the work of J. Rosenbaum, Yale
University, New Haven, CT. C, Courtesy Joel Rosenbaum, Yale University, New Haven, CT.)

Some organisms regenerate flagella if they are severed
from the cell (Fig. 38.18AB). Absence of the flagellum
activates expression of genes required to supply subunits
for regrowth of the axoneme. In approximately 1 hour,
the cell regrows a replacement flagellum, and the genes
are turned off. Even more remarkably, if only one of the
two flagella is lost, the remaining flagellum shortens
rapidly to provide components required to make two
half-length flagella (Fig. 38.18B). Then protein synthesis
slowly provides additional subunits to restore both fla-
gella to full length.
Axonemes grow at their tips by incorporation of
subunits synthesized in the cytoplasm (Fig. 38.18A). A
process called intraflagellar transport (IFT) (Fig.
38.18CD) carries individual proteins and subassemblies
such as radial spokes to the growing tip. These cytoplas-
mic cargo proteins bind one of two IFT complexes,
which associate 1:1 to form larger trains visible of
electron microscopy (Fig. 38.18C). Kinesin-2 moves IFT
trains toward the tip of the axoneme along the outer
doublets just beneath the plasma membrane. Cytoplas-
mic dynein 1b transports particles back toward the cell
body. A separate complex, called a BBSome, associates
with IFT trains and transports transmembrane proteins
(including signaling receptors) bidirectionally along
microtubules of the underlying axoneme. Movements of
transmembrane proteins allows Chlamydomonas to
glide on surfaces including the flagellum of a partner cell
during mating. Because this motion does not require
beating of the axoneme, the mechanism may represent
an early stage in the evolution of flagella.
IFT is remarkably similar to fast axonal transport (see
Figs. 37.1 and 37.3) but on a smaller scale. Cargo proteins
are loaded onto IFT complexes at the base of the cilium
and then must pass through filters located just above the
basal body. These filters have a cutoff of approximately
50kD but pass much larger IFT complexes. Proteins,
including the Ran GTPase, importins, and proteins of the
nuclear pore complex (see Chapter 9), participate in this
filtration system. After transport cargo proteins dissoci-
ate at the flagellar tip. Phosphorylation of kinesin at the
tip of the axoneme may reverse transport for the return
trip to the cell body. Trains are full of tubulin and other
axonemal proteins in growing cilia; they keep moving

bidirectionally but are largely empty when cilia are not
growing.
Rotary Cilia
Single cilia on the epithelial cells of the ventral node
of vertebrate embryos are required for the asymmetric
location of some internal organs, such as the heart and
liver, on opposite sides of the body. These nodal cilia
lack the central pair microtubules and radial spokes.
Rather than beating, the activity of the dynein arms
causes the tip of the cilia to rotate clockwise. This rotary
motion propels the extracellular fluid carrying certain
growth factors toward the left side of the embryo. The
absence of this flow explains why patients with Karta-
gener syndrome and mice missing a single dynein heavy
chain have an equal chance of having their internal
organs, such as heart and liver, positioned normally or
on the opposite side, a condition called situs inversus.
Rotary cilia may provide clues about an intermediate
stage in the evolution of axonemes.
Primary Cilia
Except for blood cells, differentiated cells in vertebrate
tissues produce a single primary cilium by growth of an
axoneme from their older mother centriole (Fig. 38.19).
Golgi Flagellum
with 9 + 0
axoneme
Centriole
Golgi
FIGURE 38.19 PRIMARY CILIUM. Electron micrograph of a thin
section of a mesenchymal cell with a primary cilium assembled from
one of the two centrioles, which serves as the basal body. (From
Fawcett DW. The Cell. Philadelphia, PA: WB Saunders; 1981.)
CHAPTER 38 n Cellular Motility 665
The axonemes lack the central pair, and most lack dynein,
so they are immotile (Fig. 38.12B).
Primary cilia are sensory organelles for both chemicals
and mechanical forces. For example, odorant receptors
of nematode olfactory neurons concentrate in the mem-
branes of primary cilia. Rod and cone photoreceptors in
the eye are modified cilia with a basal body and a vestigial
axoneme (see Fig. 27.2). Primary cilia on the epithelial
cells of kidney tubules act as flow sensors, admitting Ca2+
into the cilium through mechanosensitive channels in
the plasma membrane when bent. Many receptors con-
centrate in primary cilia including those for developmen-
tal morphogens such as sonic hedgehog and Wnt, growth
factors including platelet-derived growth factor, and
hormones like somatostatin. Some ligands activate local
Ca2+ release into the cilium.
Ciliopathies
Genetic deficiencies in the assembly of cilia or IFT result
in a remarkably wide range of human disease syndromes,
known collectively as ciliopathies. The underlying
mutations are found in more than 20 genes encoding
proteins for axonemes, basal bodies, and IFT. The defects
can appear in virtually any organ, illustrating the diverse
functions of primary and motile cilia. An early example
was polycystic kidney disease, the most common cause
of kidney failure. The most frequent mutations are in
genes for a Trp-family calcium channel, but other patients
have mutations in genes for IFT proteins. Kidney epithe-
lial cells form abnormal cysts rather than tubules, perhaps
as a result of abnormal cell division. Other ciliopathy
mutations cause defects in the central and peripheral
nervous systems, olfactory neurons, ear, liver, retina,
skeleton, and reproductive organs. Patients with some
ciliopathy syndromes are obese or have extra digits.
Box 38.1 discusses exotic eukaryotic motility systems.
Bacterial Flagella
Bacteria use a reversible, high-speed, rotary motor driven
by H+ or Na+ gradients to power their flagella (Figs. 38.24
and 38.25). Bacterial flagella differ in every respect from
eukaryotic cilia and flagella. The bacterial flagellum is an
extracellular protein wire (see Fig. 5.8), not a cytoskel-
etal structure like an axoneme inside the plasma mem-
brane. Bacteria with multiple flagella are more common
than those with single flagellum. The principles derived
from studies of Escherichia coli and a few other bacteria
apply generally, although other species exhibit many
variations on this theme.
A motor, embedded in the plasma membrane, turns
the bacterial flagellum either clockwise or counterclock-
wise (viewed from the tip of the flagellum) like the
propeller of a motorboat. In contrast to a motorboat,
moving bacteria have no momentum, so they stop in a
fraction of a nanometer if the motor stops. When
666 SECTION IX n Cytoskeleton and Cellular Motility
BOX 38.1 Exotic Eukaryotic Motility Systems
In contrast to conventional motility systems used by eukary-
otic cells discussed in the main text, a few eukaryotes
evolved exotic motility systems, four examples of which are
described here.
A Preformed Actin Filament Spring
Sperm of the horseshoe crab, Limulus, use a novel acroso-
mal process to fertilize an egg (Fig. 38.20). They preassemble
a coiled bundle of actin filaments crosslinked by a protein
called scruin. This bundle is a tightly coiled spring. An
encounter with an egg stimulates rearrangement of the
crosslinks, causing the actin bundle to unwind. Uncoiling
drives the bundle through a channel in the nucleus followed
by extension of a process surrounded by plasma membrane
that literally screws its way through the egg jelly to fuse with
the egg plasma membrane.
Calcium-Sensitive Contractile Fibers
The ciliate Vorticella avoids predators by contracting a stalk
that anchors the cell to leaves or other supports (Fig. 38.21).
The contractile fibril, called a spasmoneme, contracts
faster than any muscle. Ca2+ released from tubular mem-
branes associated with the spasmoneme triggers contrac-
tions, when it binds to spasmin, a calmodulin-like protein
that forms 3-nm filaments. Ca2+ binding changes the con-
formation of spasmin and results in rapid shortening,
because many spasmin subunits are assembled in series. The
A. Limulus B respect to the substrate as the expanding pseudopod
Egg stimulates Actin bundle
secretion of acrosome extends
and uncoiling of actin acrosomal
filament bundle process
Acrosome
C
Nucleus
Coiled
bundle
of actin
filaments
Axoneme
FIGURE 38.20 LIMULUS SPERM ACROSOMAL PROCESS.
A, Uncoiling of a bundle of actin filaments extends the acrosomal
process of the sperm of the horseshoe crab, Limulus. BC, Electron
micrograph of the actin filament bundle from the acrosomal process
of Limulus and a three-dimensional reconstruction of one filament
(yellow) decorated with crosslinking proteins (green). (Based on
the work of L. Tilney, University of Pennsylvania, Philadelphia.
BC, Courtesy W. Chiu, Baylor College of Medicine, Houston, TX.)
spasmoneme relaxes when Ca2+ dissociates. Energy for
contraction is supplied indirectly when ATP-driven pumps
create a Ca2+ gradient between the lumen of the membrane
system and cytoplasm. Movement of Ca2+ down this gradient
drives contraction.
Proteins similar to spasmin are found in other ciliates,
algae, fungi, and animals, where they are called centrin or
caltractin. These calmodulin-like proteins form fibrils that
anchor centrosomes and the basal bodies of cilia and flagella.
Mutations that inactivate caltractin in algae or yeast compro-
mise the functions of the microtubule organizers (centro-
somes or spindle pole bodies; see Figs. 34.15 and 34.20)
used for mitosis.
Major Sperm Protein, an Actin Substitute in
Nematode Sperm
Nematode sperm use amoeboid movements to find an egg
rather than swimming with flagella like other sperm (Fig.
38.22). The behavior of these sperm is so similar to a small
amoeba cell that anyone would have guessed that it is based
on the assembly of actin filaments. However, actin is a minor
protein in nematode sperm. Instead, sperm pseudopods are
filled with 10-nm wide, apolar filaments assembled from
dimers of a 14-kD protein with an immunoglobulin-like fold
called major sperm protein. Proteins in the cytoplasm and
associated with the plasma membrane guide the assembly of
the filaments, which function remarkably like actin, even
though they have no bound nucleotide and no known associ-
ated motor protein. The 10-nm filaments assemble at the
leading edge of the pseudopod and remain stationary with
A C
B
FIGURE 38.21 CALCIUM-SENSITIVE CONTRACTILE FIBERS.
AB, Light micrographs of a group of vorticellid protozoa suspended
from the bottom of a leaf, taken before (A) and after (B) contraction
of their spasmonemes. C, Electron micrograph of a thin section of
contractile fibers and tubular membranes that store and release
calcium. (Courtesy W.B. Amos, MRC Laboratory of Molecular
Biology, Cambridge, United Kingdom.)
 CHAPTER 38 n Cellular Motility 667

BOX 38.1 Exotic Eukaryotic Motility Systemscontd

A B C D

E F

Cell movement

pH 7.0 pH 6.8

FIGURE 38.22 MOTILITY OF NEMATODE SPERM. A, Scanning electron micrograph of an amoeboid sperm showing the anterior
pseudopod and trailing cell body. BC, Time series of differential interference contrast light micrographs showing movement of a live sperm
by assembly of a network of fibers at the leading edge. Arrows mark the same point in the network, which is stationary with respect to the
substrate. D, Transmission electron micrograph of an extracted sperm showing the fibers. E, Atomic model of a short segment of the sperm
filaments consisting of a polymer of major sperm protein (MSP). F, Cycle of MSP assembly at the leading edge and disassembly at the
cell body. (Courtesy T. Roberts, Florida State University, Tallahassee, and M. Stewart, MRC Laboratory of Molecular Biology, Cambridge,
United Kingdom.)

advances. Filament bundles depolymerize at the interface

between the pseudopod and the spherical cell body. A pH A B C
gradient promotes assembly of major sperm protein at the
front and disassembly at the rear of the pseudopod. This
highly efficient motility system is still unknown in other
parts of the phylogenetic tree.

Axostyles, Specialized Microtubular

Organelles
Some protozoa use dynein to generate beating movements
of large arrays of cytoplasmic microtubules called axostyles
(Fig. 38.23). The mechanism seems to be similar to an
axoneme, although the organization clearly differs. Cross-
linking structures hold together sheets of singlet microtu-
bules, which slide past each other as a result of the action
of dynein motors on adjacent sheets. Coordinated beats of
the axostyle distort the whole organism, allowing it to wiggle
about.
8 m
FIGURE 38.23 MOTILE AXOSTYLE OF SACCINOBACULUS,
A PROTOZOAN PARASITE OF TERMITES. The twisting motions
of this intracellular assembly of microtubules cause the whole para-
site to twist and turn in the gut of termites. A, Polarization light
micrograph of an isolated axostyle. B, Drawing of part of the axo-
style showing the arrangement of sheets of crosslinked microtubules.
C, Transmission electron micrograph of a cross section of the
axostyle showing microtubules crosslinked into sheets with dynein
arms between the sheets. (Courtesy R. Linck, University of Minne-
sota, Minneapolis. From Woodrum D, Linck R. Structural basis
of motility in the microtubular axostyle. J Cell Biol. 1980;87:
404414, copyright The Rockefeller University Press.)
668 SECTION IX n Cytoskeleton and Cellular Motility
A Flagella rotating counterclockwise at
(60 270 Hz) form a bundle that
propels the cell
B Clockwise rotation during
tumble (100 Hz)
C Tethered cell (20 50 Hz)
D Bead on polyhook (170 Hz)
FIGURE 38.24 DIFFERENT MANIFESTATIONS OF THE ROTA-
TION OF FLAGELLA. A, If the flagella rotate counterclockwise, they
form a bundle that propels the cell forward. B, If one or more flagella
rotate clockwise, the bundle falls apart and the cell tumbles in one
place. C, If a flagellum is tethered to a surface, the bacterium rotates.
D, If the flagellar filament is replaced by an elongated hook region with
an attached bead, the bead rotates. (Modified from Schuster SD, Khan
S. The bacterial flagellar motor. Annu Rev Biophys Biomol Struct.
1994;23:509539.)
multiple flagella are present, counterclockwise rotation
forms a bundle. Four flagella propel E. coli 30ms1, a of the proton channel, because mutations in its gene
velocity of 15 cell lengths per second, equivalent to 400
miles per hour if the bacterium were the size of an
automobile. When one or more flagella reverse their
direction and rotate clockwise, the bundle flies apart,
and the cell tumbles in one place. Figs. 27.12 and 27.13
explain how chemotactic stimuli control the probability
of clockwise rotation, favoring steady runs toward nutri-
ents and allowing for more frequent tumbles to change
direction to avoid harm.
Assays for rotation of single flagella provide insights
about the mechanism of flagellar motion (Fig. 38.24).
When a flagellum is attached to a glass slide by means of
antibodies to the flagellar filament, the bacterium rotates,
providing decisive evidence for rotation of flagella.
Similarly, beads attached to short flagella are observed to
rotate. The rotational speed depends on the resistance.
The motor of a single immobilized flagellum can rotate
a whole E. coli 10 to 50 times per second, whereas in
some species unloaded motors rotate up to 1600 times
per second (100,000rpm)!
The rotary engine driving the flagellar filament is
constructed from a rotor and stator. The cylindrical
basal body on the end of the filament rotates inside the
stator, a ring of stationary proteins embedded in the
plasma membrane and anchored to the peptidoglycan
layer (Fig. 38.25). Genetic screens for motility mutants
identified all the protein components of the motor, and
their functions were defined by analysis of the behavior
of these mutants. Most of these proteins are present in
isolated basal bodies. The functional units of the stator
consist of four MotA subunits and two MotB subunits.
MotA has four hydrophobic segments that are believed
to be transmembrane helices and a substantial cytoplas-
mic domain. MotB has one transmembrane segment and
a large periplasmic domain anchored to the peptidogly-
can layer. Flagella and basal bodies assemble but are
immotile in cells lacking either of these transmembrane
proteins. If the missing protein is replaced by initiating
its biosynthesis, the paralyzed flagellum begins to turn,
increasing its speed of rotation in a stepwise fashion, as
10 to 12 independent, torque-producing MotA4MotB2
units are added one after another.
The energy to turn the motor comes from protons
(or, in some bacteria, Na+ ions) that move down an
electrochemical gradient from outside the bacterium
through the MotA4MotB2 units to the cytoplasm. Transfer
of one proton across the membrane provides approxi-
mately the same energy as the hydrolysis of an ATP.
Pumps driven by light, oxidation, or ATP hydrolysis (see
Table 14.1) generate the proton gradient. MotA is part
inhibit both flagellar rotation and proton permeability.
The MotB transmembrane helix has a conserved aspartic
acid residue that interacts with the proton crossing
the membrane.
The mechanism producing rotation is still under
investigation, but it involves interaction of the cytoplas-
mic domain of MotA with FilG subunits on the top of the
C-ring. The transfer of a proton across the plasma mem-
brane results in a conformational change in MotA that
moves the C-ring. Roughly 1000 protons cross the mem-
brane for each rotation, corresponding to two protons
for each tiny rotational step. Proton transfer is tightly
coupled to rotation of the basal body, and the efficiency
is near 100%.
 CHAPTER 38 n Cellular Motility 669

Cap Up to 2500 nm

Filament

Hook Stator complex

D MotB

L-ring

B C P-ring
Outer membrane
Peptidoglycan
MS-ring
motor MotB MotA
Cytoplasmic membrane
MotA
Cytoplasmic E
structure:
Fli G MotA
C-ring switch complex Fli N
A Fli M

FIGURE 38.25 BACTERIAL ROTARY MOTOR. A, Drawing of the flagellar filament and rotary motor. BC, Three-dimensional reconstruction
and schematic cross section of basal body of the flagellar motor from Escherichia coli. D, Details of the stator, with schematic diagrams of the
transmembrane and cytoplasmic parts of MotA4MotB2 hexamer. E, Electron micrograph of a freeze-fractured bacterium illustrating the ring of
intramembranous particles comprising the stator of MotA4 MotB2 hexamers. (A, Modified from Schuster SD, Khan S. The bacterial flagellar motor.
Annu Rev Biophys Biomol Struct. 1994;23:509539. BC, From Zhao X, Norris SJ, Liu J. Molecular architecture of the bacterial flagellar motor
in cells. Biochemistry. 2014;53:4323433. D, Stator unit with a ribbon diagram of the C-terminal domain of MotB. Schematic diagrams of MotA
and MotB based on Kojima S, Imada K, Sakuma M, etal. Stator assembly and activation mechanism of the flagellar motor by the periplasmic
region of MotB. Mol Microbiol. 2009;73:710718. E, Courtesy S. Khan, Albert Einstein College of Medicine, New York, NY.)

SELECTED READINGS
Blanchoin L, Boujemaa-Paterski R, Sykes C, etal. Actin dynamics,
architecture, and mechanics in cell motility. Physiol Rev. 2014;94:
235-263.
Carmeliet P, Tessier-Lavigne M. Common mechanisms of nerve and
blood vessel wiring. Nature. 2005;436:193-2000.
Condeelis J, Singer RH, Segall JE. The great escape: When cancer cells
hijack the genes for chemotaxis and motility. Annu Rev Cell Dev
Biol. 2005;21:695-718.
Daniels DR. Effect of capping protein on a growing filopodium.
Biophys J. 2010;98:1139-1148.
Devreotes P, Horwitz AR. Signaling networks that regulate cell migra-
tion. Cold Spring Harb Perspect Biol. 2015;3:a005959.
Gambardella L, Vermeren S. Molecular players in neutrophil
chemotaxisfocus on PI3K and small GTPases. J Leukoc Biol.
2013;94:603-612.
Goetz SC, Anderson KV. The primary cilium: a signalling centre during
vertebrate development. Nat Rev Genet. 2010;11:331-344.
Kim S, Dynlacht BD. Assembling a primary cilium. Curr Opin Cell Biol.
2013;25:506-511.
Kolodkin AL, Tessier-Lavigne M. Mechanisms and molecules of neuro-
nal wiring: a primer. Cold Spring Harb Perspect Biol. 2011;3:a001727.
Lechtreck KF. IFT-cargo interactions and protein transport in cilia.
Trends Biochem Sci. 2015;40:765-778.
Levin M. Left-right asymmetry in embryonic development: A compre-
hensive review. Mech Dev. 2005;122:3-25.
Lin J, Okada K, Raytchev M, etal. Structural mechanism of the dynein
power stroke. Nat Cell Biol. 2014;16:479-485.
Martin AC, Goldstein B. Apical constriction: themes and variations on
a cellular mechanism driving morphogenesis. Development. 2014;
141:1987-1998.
Mizuno N, Taschner M, Engel BD, etal. Structural studies of ciliary
components. J Mol Biol. 2012;422:163-180.
Mogilner A, Rubinstein B. The physics of filopodial protrusion. Biophys
J. 2005;89:782-795.
Moriyama Y, Okamoto H, Asai H. Rubber-like elasticity and volume
changes in the isolated spasmoneme of giant Zoothamnium sp.
under Ca2+-induced contraction. Biophys J. 1999;76:993-1000.
Petrie RJ, Yamada KM. Fibroblasts lead the way: a unified view of 3D
cell motility. Trends Cell Biol. 2015;25:666-674.
Pigino G, Ishikawa T. Axonemal radial spokes: 3D structure, function
and assembly. Bioarchitecture. 2012;2:50-58.
Pollard TD, Borisy GG. Cellular motility driven by assembly and disas-
sembly of actin filaments. Cell. 2003;112:453-465.
Reiter JF, Blacque OE, Leroux MR. The base of the cilium: roles for
transition fibres and the transition zone in ciliary formation, mainte-
nance and compartmentalization. EMBO Rep. 2012;13:608-618.
Ridge KD. Algal rhodopsins: Phototaxis receptors found at last. Curr
Biol. 2002;12:R588-R590.
Ridley AJ, Schwartz MA, Burridge K, etal. Cell migration: Integrating
signals from front to back. Science. 2003;302:1704-1709.
Rrth P. Reach out and touch someone. Science. 2014;343:848-849.
Smith HE. Nematode sperm motility. WormBook. 2014;4:1-15.
Witman G. Chlamydomonas phototaxis. Trends Cell Biol. 1993;3:
403-408.
Zhao X, Norris SJ, Liu J. Molecular architecture of the bacterial flagellar
motor in cells. Biochemistry. 2014;53:4323-4433.