Cytoskeleton

-Network of protein filaments extending throughout

the cytoplasm of all eukaryotic cells.

Functions
-Providing structural support to the cell
-Provides anchorage for organelles
-Dismantles in one spot and reassembles in another to
change cell shape
-The cytoskeleton organizes the structures and activities of
the cell.
-Responsible for cell movements (entire cells or internal 1
Three main types of fibers in the cytoskeleton:
1. Microfilaments (actin filaments)
2. Intermediate filaments
3. Microtubules (tubulins)
Actin filaments
• Microfilaments, the thinnest class of the cytoskeletal
fibers, are solid rods of the globular protein actin.
– An actin microfilament consists of a twisted double
chain of actin subunits.
• Microfilaments are designed to resist tension.
• With other proteins, they form a three-dimensional network
just inside the plasma membrane. 2
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Actin exists in cells as bundles and networks:
actin-binding proteins crosslink the filaments

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 Assembly of microfilaments

Monomers of G (globular) actin polymerize to form long

filaments, the F (filamentous) actin. The addition of actin
monomers to the ends of the filament occurs at different
rates: addition to the + end, also called the barbed end
occurs 5-10x faster than to the minus or pointed end.
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Polymerization requires hydrolysis of an ATP.
All monomers are oriented in same direction, so actin
filaments have a distinct polarity: Barbed (+) & pointed (-)
ends.
Actin monomers to which ATP is bound polymerize more
readily. 7
Polymerization is reversible.
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- Actin filaments are stabilized by
filament-stabilizing proteins that
bind along their length.
- Both barbed & pointed ends can
be capped.
- Filaments can be cross-linked.
- Filaments can be split by filament-
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severing proteins.
ADF = Actin depolymerizing factor)

- ADF/cofilin bind actin filaments & increase rate of

dissociation of actin monomers from the pointed end.

- It remains bound to actin/ADP monomers, preventing their

reassembly into filaments.

- Profilin stimulates the exchange of bound ADP for ATP, so

that monomers can repolymerize.

- ADF/cofilin can also bind to & sever actin filaments.

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 Reversible polymerization of actin:
Treadmilling of actin monomers
(In some cells, remodeling of actin filaments is
responsible for half of the ATP hydrolysis in the cell)

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Growth of a microfilament requires an initial nucleation by an
actin trimer, and can stop if the rate constant for adding
monomers at the + end and one for removing them at the –
end approach the same value – at that point, treadmilling
occurs.

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Association of actin filaments with the membrane
• A network of actin
filaments & other
cytoskeletal
proteins underlies
the pl. membrane
& determines cell
shape.

• Actin bundles also

attach to the pl.
membrane to
anchor cell-cell &
cell-substratum
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contacts.
Structure of microvilli

Microvilli are relatively

stable projections of the
plasma membrane that are
supported by
microfilaments.

Microvilli increase the

surface area of cells such
as those lining the
intestine. 15
Actin filaments along with
myosin are responsible for
many types of cell
movements.
Myosin acts as a motor that
converts chemical energy
(ATP) to mechanical energy,
so generates force &
movement e.g muscle
contraction & cell division.

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• In muscle cells, thousands of actin filaments are
arranged parallel to one another.
• Thicker filaments, composed of a motor protein,
myosin, interdigitate with the thinner actin fibers.
– Myosin molecules walk along the actin filament,
pulling stacks of actin fibers together and
shortening the cell.

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Muscle contraction
Results from interaction of actin & myosin filaments. They
slide past one another.
Myosin II is present in muscle. Myosin I serve to transport
membrane vesicles & organelles along actin filaments.

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 Intermediate filaments
• 8-11 nm in diameter.
• Formed by a large, heterogeneous group of proteins
At least 4 major filament types
– Keratin - epithelial cells
– Neurofilaments – neurons
– Vimentin-containing filaments – fibroblasts, muscle
cells
– Nuclear lamina – all nucleated cells
• No energy is required for filament assembly
• Filaments are not polarized 20
Intermediate filaments in kidney cell (left, green) and cultured
epithelial cell (right, orange; desmosomes are green)

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Intermediate filaments: basic structure & assembly

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 Microtubule-based cytoskeleton
• 25 nm in diameter
• Microtubules are polymers of tubulin
• Microtubules are dynamically unstable – they can be
assembled and disassembled rapidly
• Tubules are organized by the centrosome complex
• Functions:
– Vesicle and organelle transport
– Cilia and flagella
– Mitotic spindles

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 Microtubules: α,β,γ tubulin
• Microtubules are stiff, cylindrical polymers of α and β
tubulin.
• The polymers project from the centrosome, where the γ
tubulin is present that is necessary for the “nucleation” of
the polymers.
• There is a + end projecting into the cytoplasm and a – end
that often remains anchored in the centrosome. (cilia and
flagella are a special case, as we will see).

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• Microtubules, the thickest fibers, are hollow rods.
– Microtubule fibers are constructed of the globular
protein, tubulin, and they grow or shrink as more
tubulin molecules are added or removed.
• They move chromosomes during cell division.
• Another function is
as tracks that guide
motor proteins
carrying organelles
to their destination.

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• In many cells, microtubules grow out from a
centrosome near the nucleus.
– These microtubules resist compression to the
cell.

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Formation of microtubules

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Regulation of growth & dissolution by subunit supply:
high & low concentrations of GTP-tubulin

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 Like microfilaments, microtubules in the cytoplasm exhibit
dynamic instability, including treadmilling and motility that
results from the pattern of growth, such as growing towards
and then moving chromosomes in mitosis.

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• In animal cells, the centrosome has a pair of centrioles,
each with nine triplets of microtubules arranged in a
ring.
• During cell division the centrioles replicate.

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• Microtubules are the central structural supports in
cilia and flagella.
– Both can move unicellular and small multicellular
organisms by propelling water past the organism.
– If these structures are anchored in a large structure,
they move fluid over a surface.
• For example, cilia sweep mucus carrying trapped
debris from the lungs.

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• Cilia usually occur in large numbers on the
cell surface.
– They are about 0.25 microns in diameter and 2-
20 microns long.
• There are usually just one or a few flagella
per cell.
– Flagella are the same width as cilia, but 10-200
microns long.

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• A flagellum has an undulatory movement.
– Force is generated parallel to the flagellum’s
axis.

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• Cilia move more like oars with alternating power
and recovery strokes.
– They generate force perpendicular to the cilia’s
axis.

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• In spite of their differences, both cilia and flagella have the
same ultrastructure.
– Both have a core of microtubules sheathed by the
plasma membrane (axomeme).
– Nine doublets of microtubules arranged around a pair at
the center, the “9 + 2” pattern.
– Flexible “wheels” of proteins connect outer doublets to
each other and to the core.
– The outer doublets are also connected by motor
proteins.
– The cilium or flagellum is anchored in the cell by a basal
body, whose structure is identical to a centriole. 35
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• The bending of cilia and flagella is driven by the
arms of a motor protein, dynein.
– Addition to dynein of a
phosphate group from
ATP and its removal
causes conformation
changes in the protein.
– Dynein arms alternately
grab, move, and release
the outer microtubules.
– Protein cross-links limit
sliding and the force is
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expressed as bending.
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Stem cell differentiation Maintain stem cell population

Increase stem cells

Increase differentiated cells

stem cell
differentiated cell 39
Formation of blood cells

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