CYTOSCLETONS AND

MOVEMENT OF THE CELL:

MICROFILAMENTS, INTERMEDIATE
FILAMENTS AND MICROTUBULES
ISRA WAHID
UNIT PENELITIAN
FAKULTAS KEDOKTERAN
UNIVERSITAS HASANUDDIN

Motile Cells
While most cells in the body are fixed in place by
attachments to each other and basement membranes,
some, like neutrophils and macrophages remain motile.
Free living single cells are generally motile and cell
movement plays an important in early embryogenesis.
Microfilaments and microtubules interact to control cell
movement.

The Cytoskeleton
unique to eukaryotic cells
a dynamic three-dimensional structure
that fills the cytoplasm
acts as both muscle and skeleton, for
movement and stability
the long fibers of the cytoskeleton are
polymers of subunits

Cytoskeletal Components
Actin
Microfilaments

Microtubules

Intermediate
Filaments

Idown et al. The Histochemical Journal (2000) 32:165

Scale bar represents 5m

Microfilaments
Microfilaments are fine, thread-like protein fibers,
3-6 nm in diameter.
Formed by polymerization of actin molecules (the
most abundant cellular protein)
Break down and reform rapidly, a contractile
protein.
Microfilaments' association with the protein
myosin is responsible for muscle contraction.
Microfilaments can also carry out cellular
movements including gliding, contraction, and
cytokinesis.

Microtubules
Microtubules are cylindrical tubes, 20-25 nm in
diameter.
Composed of subunits of the protein tubulin (alpha
& beta).
as a scaffold to determine cell shape, also break
down and reform readily
provide a set of "tracks" for cell organelles and
vesicles to move on
form the spindle fibers for separating chromosomes
during mitosis.
When arranged in geometric patterns inside flagella
and cilia, they are used for locomotion.

Intermediate Filaments
Intermediate filaments are about 10 nm
diameter
Formed by polymerization of proteins such as
keratin.
These were thought to be very stable but is not
always the case

provide tensile strength for the cell.

Examples of the cytoskeleton in

epithelial cells

In the intestine epithelial, all three types of fibers are present.

Microfilaments project into the villi, giving shape to the cell
surface. Microtubules grow out of the centrosome to the cell
periphery. Intermediate filaments connect adjacent cells
through desmosomes.

Actin Microfilaments
Thin, flexible filaments ~7nm
in diameter
Highly dynamic
Present in a 3-D gel
throughout the cytoplasm
Organized by over 60
accessory proteins
Primarily concentrated in
structures such as stress
fibers and cytoskeletal cortex
Diagrams from Alberts (2002) Mol Biol Cell

Microfilaments
1)

Form the cortical cytoskeleton which lies under, and is

attached to the plasma membrane and is involved in
control of cell shape

2)

Assist in forming the terminal web and the microvilli of

epithelial cells

3)

Cause movement of cells

4) Form bundles which form the contractile elements skeletal,

cardiac and smooth muscle cells

Organisation of actin

Actin microfilaments are normally found as

bundles. These may be networks as in the cortical
cytoskeleton and in smooth muscles cells, tight,
highly parallel bundles as in filopodia, microvilli
and skeletal muscle or as looser bundles as in
stress fibres

Actin controls cell shape

The cortical cytoskeleton (actin plus associated

proteins) not only determines the cell shape in
fixed cells but also changes on shape as is the
case with the platelet shown above

Function of Microfilaments
Regulation of membrane movement
Prominent in growth cones (Actin)
Dynamic changes in dendritic spine morphology

Muscle Contraction
In skeletal muscle (actin interacting with
myosin)

Local trafficking
Sensitive to local neuronal environment

Microfilaments
Abundant in
Presynaptic terminals
Dendritic spines
Growth cones

Present throughout cytoplasm

Actin cytoskelaton is universal in eukaryotes

Microfilaments
Two twisted strands of actin subunits
4-6 nm diameter
20-50 nm length (quite variable)

Microfilaments
Multiple actin genes
-actin

Four genes for four muscle types

-actin, -actin

Abundant in nervous tissue

All proteins similar (highly conserved)

Microfilaments
Proteins associated with Microfilaments
Molecular motors (eg. myosin)
Monomer actin-binding proteins

Regulate amount of actin assembled into

microfilaments by sequestering actin monomers

Capping proteins

Anchor microfilaments to other structures

Regulate microfilament length
Mutation in Schwann cells causes neurofibromatosis 2

Microtubules
Thick, rigid tube ~25 nm in
diameter
Highly dynamic
Present throughout the
cytoplasm
Higher order structures are
not observed

Diagrams from Alberts (2002) Mol Biol Cell

Microtubules
1) Organise the endoplasmic
reticulum and the Golgi
apparatus
2) Act as a railroad connecting
the trans golgi network to the
cell surface and the early
endosome compartments to
the late ones
3) Form the spindle apparatus in
mitotic cells
4) Act as motile elements in cilia
and flagella

Function of Microtubules
Cell movement
Functional core of cilia and flagella

Mitotic spindle
Organelle involved in cell division

Inhabitants of axons and dendrites

Intracellular transport
Essential for fast-axonal transport

Cell Morphology

Microtubules
Smallest subunit is tubulin

10% of total brain protein

and tubulin
50 kDa proteins
Multiple genes for both types
Different gene products are enriched or
specific to neurons
Different gene products are expressed at
specific times in development

Microtubules
Second smallest subunit is "gobule"
Heterodimer of and tubulin

Protofilaments
Linear arrangement of globular subunits

12-14 protofilaments form microtubule

25 nm diameter, hollow tube
Up to hundreds m length
Polarized: +(fast) and (slow-growing) ends

Microtubules

Intermediate Filaments
Semi-flexible filaments
~10nm in diameter
Very stable proteins
Forms a 3-D gel throughout
the cytoplasm
Protects the cell from
overloading

Diagrams from Alberts (2002) Mol Biol Cell

Intermediate Filaments
Also called Neurofilaments
Five classes
Type I, II: Keratin (hair and nails)
Type V: nuclear lamins
Type III, IV: neuronal function

Intermediate Filaments
1) Form cables which stretch across the cell from
desmosomes on one side to desmosomes on
the other so giving strength to the cells
2) Hold the nucleus in place
3) May play a role in organising permanent cell
extensions such as nerve axons
4) The nuclear lamina is formed by proteins called
lamins whichare closelyrelated to intermediate
filament proteins

Intermediate
filaments

-concentrated in
the region round
the nucleus and
holding the
nucleus in place
Others raddiate to
the cell surface
and attach to
desmosomes /
hemi-desmosomes
giving strength to
the cell.

Reinforced tissue
Distribution of the cytokeratin
filaments (green) of cultured
epithelial cells as compared
with the plasma membrane
(blue).

Desmosome bind both stains

and appear pale blue.

Molecular Motors
Molecules that hydrolyze ATP (ATPase)
Drive cell movement such as axonal
transport
Three types
Myosin

Muscle contraction via interaction with

microfilaments

Dynein
Kinesin

Cellular Motors

Kinesin
Responsible for fast axonal transport
toward distal (terminal) end
Head attaches to microtubule
Tail attaches to organelle
Hydrolysis of ATP moves kinesin head distally,
toward plus end of microtubule

Strongly inhibited by adenylylimidodiphosphate (AMP-PNP;

nonhydrolyzable ATP analog)

Kinesin
Some kinesins are
Head

Tail

Monomers (KIF1A)
Trimers (KIF3A/B)

Head contains ATP

binding and
microtubule
binding domains

Dynein
Microtubule transport
Anterograde direction

Substrate is actin filaments or long microtubules

Retrograde direction

Substrate is long microtubules

MAP1c is one type

40 nm in length

Weakly inhibited by AMP-PNP

Dynein Structure

Myosins
First identified in skeletal muscles
Myosin I, II and V found in nervous
system
Myosin VI and VIIA also in nervous
system
Implicated incongenital deafness

Likely role in growth cone motility

Myosin Structure

Myosin I
Structure
Single heavy chain

Function
Interacts directly with membrane surfaces
May generate movement of plasma membrane
components
Mechanotransduction (myosin IB expressed in
stereocilia of hair cells)

Myosin II
Structure
Dimer composed of two heavy chains
Two dimers may form bipolar filaments

Function
Contractile ring in mitosis
Unknown role in neurons

Myosin V
Structure
Dimer composed of two heavy chains
Multiple calmodulin binding sites

Function
Found in growth cones
"Dilute" mutation results in seizures in adult
mice