CYTOSKELETON AND CELL MOTILITY

Cell biology and its application

BI-1202

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 Cytoskeleton is structural protein that builds up membrane system and
cytoplasmic components
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 Summary schematic showing locations of microtubule and actin filament populations in a
large paused growth cone and developing axon branch.

Dent E W , Kalil K J. Neurosci. 2001;21:9757-9769

©2001 by Society for Neuroscience

3 TYPES OF
CYTOSKELETON:
• Microtubules determine
the positions of
membrane-enclosed
organelles and direct
intracellular transport
• Actin filaments
determine the shape of
the cell's surface and are
necessary for whole-cell
locomotion
• Intermediate filaments
provide mechanical
strength and resistance
to shear stress.

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 Cytoskeleton functions:
1. Determine cell shape and cell
defense
2. Placement of organelles in the
cell
3. Materials and organelles
movement in the cell
4. Cell movement
5. Place for anchoring mRNA
and help the translation of
mRNA become polypeptide
6. Important component in cell
division
IF (4-5) = cell adhesion and support
ACTIN (1-4) = maintains cell shape and generates force for movement

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MT: thick pipes
 MT: monorails

Vesicle
ATP
Receptor for
motor protein

Motor protein Microtubule

(ATP powered) of cytoskeleton
Kinesin or Dynein
MF: smallest of the three
 Action/Myosin Fibers: muscle
contraction

Muscle cell
Actin filament

Myosin filament
Myosin arm

Myosin motors in muscle cell contraction

IF: the least dynamic
• Monomer-polimer :
1. Microtubule : Tubulin
2. Filament : Actin
3. Intermediate Filamen :
“fibrous”protein

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 MICROTUBULE
Structure and composition :
• Rod shape (tubule)
exist almost in all eukaryote cells
• Function in mitosis and cell movement
cilia dan flagella
• Consist of proto -filament  paralel
along the axis of tubules.
• Protofilament consists of 2 kind of
tubulin molecules : α dan 

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 Bind to MAP (microtubule associated protein)

Substances that interfere microtubule :

- nokodazol  inhibit polimerisation 
substance binds to tubulin inhibit + end
addition
- colchicine  de-polimerisation
• from the Autumn Crocus (a lavender)
• causes disassembly of microtubules

- vinblastin & vincristin  depolimerisation of

microtubules
- taxol  increase microtubule’s stability
Other functions:
• Mechanical – physical + give shape
• Neuron: vesicles pathway and cytoplasmic particles
• In embryo : conserve elongated shape from axon
• conserved internal organization of the cell
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Microtubule play a role in intracellular
motility : material transport
Protein motor : myosin, kynesin,
dynein

myosin – aktin
kynesin  vesicle movement from cell
to synaptic terminal & organelles
transport
2012 mit/rre/egr,SITH dynein  cilia and flagella movement
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 Cooperation of
General model of kynesin and dynein myosin and kynesin
mediated transport at the cell cortex

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Microtubule-Organizing center (MTOC): the place where enucleation
happen
→ forming molecule start to organize and elongation also started
→ centrosome, basal body
Centrosome
- Only in animal cells
- Consists of 2 centrioles & peri-centriolar material

- Microtubules from cilia and flagella start from basal body

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Organization of MT around the MTOC and spindle poles
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• In most animal cells, there
is a single, well-defined
MTOC called the
centrosome, located near
the nucleus. From this focal
point, the cytoplasmic
microtubules emanate in a
star-like, “astral”
conformation.
• A centrosome is composed
of a fibrous centrosome
matrix
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Centrosome.
(A) The centrosome is the major MTOC of animal
cells. Located in the cytoplasm next to the
nucleus, it consists of an amorphous matrix of
protein containing the γ-tubulin ring complexes
that nucleate microtubule growth. This matrix is
organized by a pair of centrioles, as described in
the text.
(B) A centrosome with attached microtubules. The
minus end of each microtubule is embedded in
the centrosome, having grown from a γ-tubulin
ring complex, whereas the plus end of each
microtubule is free in the cytoplasm.
 Example : axon transport
- Cell body : nucleus, ER, Golgi  synthesis protein
- Axon  neurotransmitter transport by microtubules anterograde
Microtubues movement by protein motor  produce energy

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 Cilia and Flagella

• Microtubules control the beating of cilia and flagella,

locomotor appendages of some cells
Cilia and Flagella
 Cilia and Flagella
• Cilia and flagella share a common ultrastructure:
– Flagella is longer than cilia
– Consists of axonem (center) that surrounded by 9
double microtubules
– MTOC : basal body  A basal body that anchors
the cilium or flagellum
– A motor protein called dynein, which drives the
bending movements of a cilium or flagellum
 0.1 µm Outer microtubule Plasma
doublet membrane
Dynein arms
Central
microtubule
Cross-linking
Microtubules proteins inside
outer doublets
Plasma
membrane Radial
Basal body spoke

0.5 µm 0.1 µm
Triplet

Cross section of basal body

The arrangement of microtubules in a flagellum or cilium.
• (A) Electron micrograph of the flagellum of a green-alga
cell (Chlamydomonas) shown in cross section,
illustrating the distinctive “9 + 2” arrangement of
microtubules.
• (B) Diagram of the parts of a flagellum or cilium. The
various projections from the microtubules link the
microtubules together and occur at regular intervals
along
2012 the length of the axoneme.
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 The contrasting motions of flagella and cilia.
(A)The wave-like motion of the flagellum of a
sperm cell from a tunicate. The cell was
photographed with stroboscopic
illumination at 400 flashes per second. Note
that waves of constant amplitude move
continuously from the base to the tip of the
flagellum.
(B)The beat of a cilium, which resembles the
breast stroke in swimming. A fast power
stroke (red arrows), in which fluid is driven
over the surface of the cell, is followed by a
slow recovery stroke. Each cycle typically
requires 0.1–0.2 sec and generates a force
perpendicular to the axis of the axoneme

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 Actin Filaments
• The tip of the leading edge
of a cell nucleates actin
filaments.

• Actin filament nucleation

most frequently occurs at
the plasma membrane 
highest density of actin
filament is at the cell
periphery

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 Cell movement: Actin
Actin polymerization produce
Cell movement
Cell shape

Movement is helped by myosin

- vesicles movement (in actin filaments)
- “Contractile bundles” (eg:cytokinesis)
- “Thick and thin filaments” (eg.muscle
contraction )

Cell movement:
“membrane extension, cell-substrate
adhesion, cell body translocation,
breaking cell attachments”

2. Outside signal induces cell migration

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 Microfilament

Functions:
Motility, cell shape and
cytokinesis
A.Anti-parallel bundles+myosin = contraction (EX: cytokinesis, cell migration)
•Consist of actin protein (,) B.2D gel = structural support
& tropomyosin  filament C.Parallel fibers = protrusions

•Interaction with myosin

assembly and disassembly  C
depends on ATP B
A
•Substance taht can be
depolimerised:cytokalasin,

•phalloidin increase stability of

microfilament

2012 mit/rre/egr,SITH Actin binding proteins create different actin arrays in a crawling cell
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 Not-so-mellow mushrooms
• Phallotoxin (phalloidin)
– an actin filament stabilizer
– the poison in some mushroom genera
– It kills by stabilizing actin
filaments (inhibiting disassembly)
– Immediate cause of death is
liver failure
• Cytochalasin
– an actin filament de-stabilizer
– also derived from mushrooms
Death Cup mushroom
Motor proteins transition/cycle between different conformations:
one step is driven by the hydrolysis of ATP,
thereby making the cycle essentially irreversible and movement unidirectional

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Molecular basis of cell crawling
 Biconcave Red Blood Cells
Why are erythrocytes good for plasma
membrane and cortical cytoskeleton
studies?
the structural basis for the cortical
cytoskeleton in erythrocytes-spectrin
 Wound healing in blood
vessel

Injury to lining of blood

Platelet plug forms Fibrin clot traps
vessel exposes connective blood cells
tissue; platelets adhere

Platelet releases chemicals

that make nearby platelets sticky

Platelets Clotting factors from:

Damaged cells

Prothrombin Thrombin
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Some intracellular pathogens such as the bacteria Listeria and Shigella and the vaccinia
virus usurp the host cell’s mechanism of assembling actin networks and propel
themselves through the cytoplasm with actin “tails”.
Listeria lifecycle: an intracellular
pathogen
• Phagocytosis
• Lysis of phagosome
• Proliferation in the
cytoplasm
• Locomotion
• Cell-cell spread
through filopodia

• How does Listeria

accomplish cell-cell
spread? http://www.med.ufl.edu/biochem/DLPURICH/Listeria.html
 Stealing the machinery
• Listeria has on its surface the protein ActA
• ActA recruits Arp2/3 from the cytoplasm and
activates it (basically substituting for WASP)
• Promotes actin filament nucleation and growth

Listeria
Intermediate filament

Resistant to pressure, e.g in cornified

skin (including human skin) IF , skin
is waterproof, resistant against bacteria
or chemical substances

Assembly and disassembly  because

phosphorilasi dan defosforilasi subunit

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Mutant of keratin gene causes peel
of/ wound in the skin
(Epidermolysis bullosa simplex)

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