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
(ATP powered)

Kinesin or Dynein

Microtubule
of cytoskeleton

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


1.
2.
3.

Monomer-polimer :
Microtubule : Tubulin
Filament : Actin
Intermediate Filamen :
fibrousprotein

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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 : 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 microtubules 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
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dynein cilia and flagella movement

General model of kynesin and dynein

mediated transport

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Cooperation of
myosin and kynesin
at the cell cortex

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

 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 -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 -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 m

Outer microtubule
doublet
Dynein arms
Central
microtubule
Cross-linking
proteins inside
outer doublets

Microtubules
Plasma
membrane
Basal body

0.5 m

Radial
spoke

0.1 m
Triplet

Cross section of basal body

Plasma
membrane

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
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 0.10.2 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
Consist of actin protein (,)
& tropomyosin filament
Interaction with myosin
assembly and disassembly
depends on ATP

A.Anti-parallel bundles+myosin = contraction (EX: cytokinesis, cell migration)

B.2D gel = structural support
C.Parallel fibers = protrusions

C
A

Substance taht can be

depolimerised:cytokalasin,

phalloidin increase stability of

microfilament
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Actin binding proteins create different actin arrays in a crawling cell

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

vessel exposes connective
tissue; platelets adhere

Platelet plug forms Fibrin clot traps

blood cells

Platelet releases chemicals

that make nearby platelets sticky

Platelets Clotting factors from:

Damaged cells
Prothrombin
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Thrombin

Fibrinogen

Fibrin

Some intracellular pathogens such as the bacteria Listeria and Shigella and the vaccinia
virus usurp the host cells 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
http://www.med.ufl.edu/biochem/DLPURICH/Listeria.html
spread?

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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