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 Intermediate
Microfilaments Microtubules Filaments

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

Scale bar represents 5m
 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
-concentrated in the
filaments 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 adenylyl-
imidodiphosphate (AMP-PNP;
nonhydrolyzable ATP analog)
 Kinesin
• Some kinesins are
– Monomers (KIF1A)
– Trimers (KIF3A/B)
Head Tail
• 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