CHAPTER

9
The Cytoskeleton and Cell Motility
Required reading: chapter 9 appropriate sections

Remember to use the all the

on-line resources; the
glossery, flashcards quizzes,
practice questions (under
assignments) and review the
following animation – “
9.1 | Overview of the Major Functions of the Cytoskeleton
Properties of cytoskeletal components
9.1 | Overview of the Major Functions of the Cytoskeleton
Properties of cytoskeletal components
9.1 | Overview of the Major Functions of the Cytoskeleton
Roles of cytoskeletal components
9.1 | Overview of the Major Functions of the Cytoskeleton
Roles of cytoskeletal components
 9.1 | Overview of the Major Functions of the Cytoskeleton
Roles of cytoskeletal components:
cytoskeletal changes during cell division

Division of a polarized fibroblast cell

Purple - Actin filaments
Green - Microtubules
Brown - DNA

Cell polarity = spatial differences in shape, structure,

and function within a cell
Almost all cell types exhibit some form of polarity,
which enables them to carry out specialized functions
Epithelial cells have apical-basal polarity
Motors and the cytoskeleton

Kinesin – Anterograde MT motor

Dynein – Retrograde MT motor
the cytoskeleton and signaling
 9.2 | Structure and Function of Microtubules
Structure and Composition of Microtubules
• Microtubules are the largest of the cytoskeletal components of a cell
• There are two types of microtubules
• They are involved in a variety of functions in the cell

1) Cytoplasmic microtubules pervade the cytosol and

are responsible for a variety of functions:

• Maintaining axons (in nerve cells)

• Formation of mitotic and meiotic spindles
• Placement and movement of vesicles
• Maintaining or altering cell shape

2) Axonemal microtubules include the organized

and stable microtubules found in structures
specialized for movement such as:

• Cilia
• Flagella
• Basal bodies to which cilia and flagella attach

The axoneme, the central shaft of a cilium or

flagellum, is a highly ordered bundle of MTs
 9.2 | Structure and Function of Microtubules
Structure and Composition of Microtubules

• MTs are straight, hollow

cylinders of varied length
that consist of (usually 13)
longitudinal arrays of
polymers called
protofilaments

• The basic subunit of a

protofilament is a
heterodimer of tubulin, one
α-tubulin and one β-tubulin
(globular proteins)

• These bind noncovalently

to form an αβ-heterodimer,
which does not normally
dissociate
 9.2 | Structure and Function of Microtubules
Microtubule-Associated Proteins
Microtubules typically contain additional
proteins, called microtubule-associated
proteins (or MAPs)

MAPs comprise a heterogeneous collection

of proteins with one domain that attaches to
the side of a microtubule and another
domain that projects outward as a tail
MAPs generally increase the stability of
microtubules and promote their assembly

MAP activity is controlled by the addition

and removal of phosphate groups from
amino acid residues
Schematic diagram of a brain
MAP2 molecule bound to the
An abnormally high level of surface of a microtubule.
phosphorylation of one particular MAP,
called tau, has been implicated in
Alzheimer’s disease
 9.2 | Structure and Function of Microtubules
Structure and Composition of Microtubules
Building block of a microtubule is a dimer of
α tubulin and β tubulin (each 55,000 Da)

α and β tubulin are two related globular

proteins (40% amino acid identity)
noncovalently bound together (they are not
found as individual monomers in the cell)

Each monomer has a GTP binding site

GTP in the α monomer is physically trapped at the dimer interface

It is never hydrolyzed or exchanged

In the β tubulin monomer the nucleotide can

be in either GDP or GTP form (and is both
hydrolyzable and exchangable)
 9.2 | Structure and Function of Microtubules
Structure and Composition of Microtubules

• All the dimers in the MT are oriented the same way

• Because of dimer orientation, protofilaments have an inherent polarity
• The two ends differ both chemically and structurally

• minus(-) end and a plus (+) end

 9.2 | Structure and Function of Microtubules
Microtubules as Structural Supports and Organizers

Microtubules resist forces that might

compress or bend the fiber for
mechanical support
Distribution of microtubules helps
determine the shape of that cell
In cultured animal cells, microtubules
extend in a radial array outward from
near the nucleus, giving cells a round,
flattened shape
Microtubules of columnar epithelial Localization of microtubules in a
cells are oriented with their long axis cultured mouse cell shown by
fluorescent anti-tubulin antibodie
parallel to the long axis to support the
cell’s elongated shape Microtubules extend from the
perinuclear region of the cell in a
radial array and curve gradually as
they conform to the shape of the cell.
 9.2 | Structure and Function of Microtubules
Microtubules as Structural Supports and Organizers
 9.2 | Structure and Function of Microtubules
Microtubules as Structural Supports and Organizers

Microtubules also play a key role in

maintaining the internal organization of cells

Treatment of cells with nocodazole or

colchicine, which promote microtubule
disassembly, can disperse the Golgi
elements into separate Golgi stacks
scattered throughout the cytoplasm

When the drug is removed and microtubules

reassemble, the Golgi membranes return to
their normal position in the cell interior

red = microtubules

Green = Golgi
 9.3 | Motor Proteins: Kinesins and Dyneins

• Motor Proteins that Traverse the

Microtubular Cytoskeleton
– Molecular motors convert
energy from ATP into
mechanical energy.
– Molecular motors move
unidirectionally along their
cytoskeletal track in a
stepwise manner.
– Three categories of molecular
motors:
• Kinesin and dynein move
along microtubule tracks.
• Myosin moves along
microfilament tracks.
 9.3 | Motor Proteins: Kinesins and Dyneins
Motor Proteins Traverse the Microtubular Cytoskeleton
Motor proteins move unidirectionally along their
cytoskeletal track in a stepwise manner and
undergo a series of conformational changes that
constitute a mechanical cycle
The steps of the mechanical cycle are coupled
to the steps of a chemical cycle, which provide
the energy necessary to fuel the motor’s activity
and to move along the track
The chemical cycle steps include the binding of
ATP to the motor, the hydrolysis of ATP, the
release of the products (ADP and Pi), and the
binding of a new molecule of ATP
Motor proteins have virtually no momentum and
are subjected to tremendous frictional
resistance from their viscous environment
As a result, a motor protein comes to a stop
almost immediately once energy input has
ceased
 9.3 | Motor Proteins: Kinesins and Dyneins
Kinesins
kinesins :the smallest and best understood microtubular motors

The kinesin molecule is in a superfamily

of 14 KRPs (kinesin-related proteins)
kinesin molecule: tetramer of two
identical heavy and two identical light
chains
The globular heads bind microtubules
and act as ATP-hydrolyzing, force-
generating engines Force-generating heads bind to
the microtubule; tail binds to the
Each head is connected to a neck, a cargo being transported
rodlike stalk, and a fan-shaped tail that
binds cargo.
A number of different proteins have been
identified as potential adaptors that link
specific KRPs and their cargoes
 9.3 | Motor Proteins: Kinesins and Dyneins
Kinesins
hand-over-hand model
In vitro motility assays show that
kinesin is a plus end-directed
microtubular motor

Each kinesin step is approximately

8 nm, the length of one tubulin
dimer, and requires the hydrolysis
of a single ATP molecule

Kinesin movement:
• is proportional to the ATP
concentration
• moves in a “hand-over-hand”
mechanism
• is highly processive (can walk
along a microtubule for
considerable distances without
falling off)
 9.3 | Motor Proteins: Kinesins and Dyneins
Kinesins

The two kinesin heads act in a

coordinated manner, so that they
are at different stages in their
chemical and mechanical cycles at
any given time
 9.3 | Motor Proteins: Kinesins and Dyneins
Kinesin-Mediated Organelle Transport
Alteration in the microtubule AB microtubule AB
phenotype of a cell
lacking a member of
the kinesin
superfamily
Control KO
Cells from a normal
9.5 day mouse or one
that lacked the gene Mito AB Mito AB
for kinesin-like protein
KIF5B

Mitochondria of the
Control KO
KIF5B-deficient cell
are absent from the
cell’s peripheral
regions.
 9.3 | Motor Proteins: Kinesins and Dyneins
Cytoplasmic Dynein
Cytoplasmic dynein is a huge protein
composed of two identical heavy chains
and a variety of intermediate and light
chains
The heavy chain consists of a large globular
force-generating head and a microtubule-
binding stalk
Cytoplasmic dynein moves processively
along a microtubule toward the polymer’s
minus end—opposite that of most kinesins
Role 1: Position the spindle and move
chromosomes during mitosis
Role 2: Position the centrosome and Golgi
complex and moving organelles, vesicles,
and particles through the cytoplasm

note that axonemal dyneins that generate motility in cilia and flagella will be covered later
9.3 | Motor Proteins: Kinesins and Dyneins
Cytoplasmic Dynein
Green = ER
Dynein-driven cargo includes: Red = microtubules
• endosomes and lysosomes
• ER-derived vesicles heading toward the
Golgi complex
• RNA molecules
• HIV virus which is transported to the
nucleus of an infected cell

Cytoplasmic dynein does not interact directly

with membrane-bounded cargo but requires the
intervening adaptor dynactin, a protein that also
increases the processivity of dynein
 9.3 | Motor Proteins: Kinesins and Dyneins
Cytoplasmic Dynein

Kinesins and cytoplasmic dynein move similar materials

in opposite directions over the same railway network

Organelles may bind kinesin and dynein simultaneously

Movement along microtubules can be regulated

Melanosome aggregation is
via dynein

Melanosome disperation
via kinesin
learning objectives:

be clear on the structure and composition of microtubules

be clear on the structure and function of kinesins and dyenins