CYTOSKELETON AND CELL MOTILITY

Cell biology and its application

BI-1202

2012 mit/rre/egr,SITH ITB

Cytoskeleton is structural protein that builds up membrane system and cytoplasmi
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.

2012 mit/rre/egr,SITH ITB

 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

Types of IF: Neurofilament neuron; keratin epithelial cells,

Cortex of nucleus -.., etc
 Monomer-polimer :
1. Microtubule : Tubulin
2. Filament : Actin
3. 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
One type of addition
MAP2 : Tau
Alzheimer - colchicine de-polimerisation
problem with tau from the Autumn Crocus (a lavender)
protein 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
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myosin aktin
kynesin vesicle movement from
cell to synaptic terminal & organelles
transport
dynein cilia and flagella
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Organization of MT around the MTOC and spindle poles
 Cooperation of myosin and
General model of kynesin and dynein mediated transport kynesin 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
- Polymerization in
MTOC is started with
the arrangement of -
tubulin in the
nucleation center and
then polymerization
will continue with the
arrangement of and
tubulin

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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 (A) The centrosome is the major MTOC of animal
conformation.
A centrosome is composed
of a fibrous centrosome
matrix
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Centrosome.
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

2012 mit/rre/egr,SITH ITB

 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
 Outer microtubule Plasma
0.1 m
doublet membrane
Dynein arms
Central
microtubule
Cross-linking
Microtubules proteins inside
outer doublets
Plasma
membrane Radial
Basal body spoke

0.5 m 0.1 m
Triplet

Cross section of basal body

 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
cortex

2012 mit/rre/egr,SITH ITB

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

1)
1) RBCs
RBCshave
haveproven
provenparticularly
particularlyuseful
usefulfor
forstudies
studiesof
ofboth
both
the plasma membrane and the cortical cytoskeleton.
the plasma membrane and the cortical cytoskeleton.
This
This is
is because
because RBCs
RBCs have
have no
no nucleus
nucleus or
or internal
internal
organelles. In addition, human erythrocyteslack
organelles. In addition, human erythrocytes lackother
other
cytoskeletal components (microtubules
cytoskeletal components (microtubules and and
intermediate
intermediatefilaments),
filaments),so sothe
thecortical
corticalcytoskeleton
cytoskeleton
isisthe
the principal determinant of their distinctiveshape
principal determinant of their distinctive shapeasas
the structural basis for the cortical
cytoskeleton in erythrocytes-spectrin
 Wound healing in blood vessel

Injury to lining of blood Fibrin clot traps

Platelet plug forms
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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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
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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