Cytoskeleton

(1st half)

 Cytoskeletonan intricate network of protein

filaments that extends throughout the cytoplasmhelp
eucaryotic cells to adopt a variety of shapes, organize
the many components in their interior, interact
mechanically with the environment, and carry out
coordinated movementsimportant in animal cells (no
cell wall). Some cytoskeletal components are present in
bacteria.
Cytoskeletona highly dynamic structure i.e.
continuously reorganized ( thus are bones & muscles
of a cell)controls the location of the organelles &
providing the machinery for the transport between them
responsible for the segregation of chromosomes into
daughter cells & pinching apart of cells
Without the cytoskeletonwounds would never heal,
muscles would be useless, & sperm would never reach
the egg.

Fig: The cytoskeleton gives a cell its shape and

allows the cell to organize its internal
components (the
microtubules (green) & the actin filaments (red)

 The cytoskeletonthree types of protein

filamentsintermediate filaments,
microtubules, & actin filamentseach
has distinct mechanical properties &
formed from a different protein subunit.
Intermediate filamentsformed by a
family of fibrous proteins.
Microtubulestubulin is subunit
Actin filamentactin is subunit
In each case, thousands of subunits
assemble into a fine thread of protein that
sometimes extends across the entire cell.

Intermediate Filaments
Intermediate filamentsgreat tensile strength
enable cells to withstand the mechanical stress that
occurs when stretchedfirst discovered in the
smooth muscle cellsdiameter (about 10 nm) is b/w
that of the thin actin-containing filaments & the
thicker myosin filamentstoughest & durable of the 3
typesthe only one survive under treatment with
concentrated salt solutions & non-ionic detergents
found in the cytoplasm of most animal cellsforming
a network throughout the cytoplasm, surrounding the
nucleus & extending out to the cell peripheryoften
anchored to the plasma membrane at cellcell
junctions such as desmosomesalso found within
the nucleus, nuclear lamina (a mesh of intermediate
filaments)strengthens the nuclear envelope.

intermediat
e
filaments

desmosom
e
connecting
two cells

Fig: Intermediate filaments form a

strong, durable network in the
cytoplasm of the cell

Intermediate Filaments are strong

and ropelike
Intermediate filamentslike ropeswith many long
strands twisted togetherits subunits are elongated
fibrous proteinseach composed of an N-terminal
globular head, a C-terminal globular tail, & a central
elongated rod domain which consists of an extended helical region (form stable dimers)Two of these coiledcoil dimers then associate by non-covalent bonding to
form a tetramer which then bind to one another end-toend and side-by-side to generate the final ropelike
intermediate filament. The central rod domains of different
intermediate filament are all similar in size & amino acid
sequencealways form filaments of similar diameter &
internal structure the globular head & tail regions, which
are exposed on the surface of the filament, allow it to
interact with other components of the cytoplasm.

Fig: Intermediate filaments are like ropes

made of long, twisted strands of protein .

Intermediate Filaments strengthen

cells against mechanical stress
Intermediate filamentsare prominent in cells
subjected to mechanical stresse.g. along the
length of nerve cell axons, in muscle cells & in
epithelial cellscan be grouped into four classes:
(1) keratin filaments in epithelial cells
(2) vimentin and vimentin-related filaments in
connective-tissue cells, muscle cells, and supporting
cells of the nervous system (glial cells);
(3) neurofilaments in nerve cells;
(4) nuclear lamins, which strengthen the nuclear
membrane of all animal cells.
The first threefound in the cytoplasm & the fourth
in the cell nucleus

Fig: Intermediate filaments can be

divided into several different
categories.

 The keratinsEvery kind of epithelium in the

vertebrate bodyhas its own distinctive
mixture of keratin proteinsSpecialized keratins
also occur in hair, feathers, & clawsformed
from a mixture of different keratin subunits
typically span the interiors of epithelial cells &
filaments are indirectly connected through
desmosomesdistributes the stress that occurs
when the skin is stretched.
In rare human genetic disease epidermolysis
bullosa simplex, mutations in the keratin
genesinterfere with the formation of keratin
filaments in the epidermisthus the skin is
highly vulnerable to mechanical injuryeven a
gentle pressure can rupture its cells.

 Many intermediate filamentsfurther stabilized &

reinforced by accessory proteinsplectinthese
proteins also link intermediate filaments to
microtubules, to actin filaments, & to adhesive
structures in the desmosomes.
Mutations in the gene for plectindisease that
combines features of epidermolysis bullosa
simplex (caused by disruption of skin keratin),
muscular dystrophy (caused by disruption of
intermediate filaments in muscle), and
neurodegeneration (caused by disruption of
neurofilaments).
Thus, plectin may not be necessary for the initial
formation of intermediate filaments, its crosslinking
action is required to provide cells with the strength
they need to withstand the mechanical stresses.

Fig: Plectin aids in the bundling of intermediate

filaments and links these filaments to other cytoskeletal
protein networks, plectin (green), intermediate filaments
(blue), microtubules (red).

The nuclear envelope Is supported by

a meshwork of Intermediate Filaments
The intermediate filamentswithin tough nuclear
laminaare constructed from laminsthe intermediate
filaments of the nuclear lamina disassemble & re-form
at each cell division (when nuclear envelope breaks
down during mitosis & then re-forms)controlled by the
phosphorylation & dephosphorylation of the
lamins. When phosphorylatedthe consequent
conformational change weakens the binding between
the tetramers & causes the filament to fall apart&
Dephosphorylationcauses reassembly.
Defects in a particular nuclear laminprogeriacause
affected individuals to appear to age prematurelyhave
wrinkled skin, lose their teeth and hair, and often
develop severe cardiovascular disease by the time they
reach their teens.

Fig: Intermediate filaments support

and strengthen the nuclear envelope.

Microtubules
Microtubuleslong & relatively stiff hollow tubes of
proteincan rapidly disassemble in one location &
reassemble in anothergrow out from a small
structure near the center of the cell called the
centrosome in an animal cellExtending out
toward the cell peripherycreate a system of tracks
within the cell, along which vesicles, organelles, and
other cell components are moved. When a cell
enters mitosismicrotubules disassemble & then
reassemble into mitotic spindlethat will
segregate the chromosomes equally into the two
daughter cellscan also form permanent structures
e.g., cilia & flagellaused as a means of propulsion
or to sweep fluid over the cell surface

Fig: Microtubules usually grow out of

an organizing structure

Microtubules are Hollow tubes with

structurally distinct ends
Microtubulestubulin, in which each
subunit a dimercomposed of -tubulin
& -tubulinThis tubelike structure is
made of 13 parallel protofilaments
which has a structural polarity with tubulin exposed at one end and -tubulin
at the otherOne end of the microtubule,
thought to be the -tubulin end, is called
its plus end, & the other, the -tubulin
end, its minus endis crucial, both for
the assembly of microtubules & for their

Fig: Microtubules are hollow

tubes of tubulin.

The centrosome Is the major microtubuleorganizing center in animal cells

Microtubulesformed by outgrowth from specialized
organizing centerse.g. in animals, the centrosome which
is close to the cell nucleus contain hundreds of ring-shaped
structures formed from another type of tubulin-tubulin
each ring serves as the starting point, or nucleation site
The -tubulin dimers add to the -tubulin ring in a specific
orientationminus end is embedded in the centrosome &
growth occurs only at the plus end i.e at the outward- facing
end. The centrosomemay also contains a pair of centrioles
(lacking in plants)made of a cylindrical array of short
microtubules. Microtubules need nucleating sites. In the
living cell, the concentration of free tubulinis too low
to drive the difficult first step of assembling the initial ring of
a new microtubule. By providing organizing centers
containing nucleation sites, and keeping the concentration of
free -tubulin dimers low, cells can thus control where
microtubules form.

Fig: Tubulin polymerizes from

nucleation sites on a centrosome

Growing microtubules show dynamic

Instability
Once a microtubulenucleatedplus end grows
outward by by the addition of -tubulin subunits
Then the microtubule suddenly undergoes a transition
causing it to shrink inward by losing subunits from
its free endstart growing again or it may disappear
completely, to be replaced by a new microtubule
behavior, known as dynamic instabilitystems from
the intrinsic capacity of tubulin molecules to hydrolyze
GTP. Each free tubulin dimercontains one tightly
bound GTP molecule that is hydrolyzed to GDP (still
tightly bound) shortly after the subunit is added to a
growing microtubule. The GTP-associated tubulin
moleculespack together in the wall of the
microtubule & those carrying GDP bind less strongly.

 When polymerization is proceedingtubulin

molecules add to the end of the microtubule faster
than the GTP they carry is hydrolyzedThe end of
a growing microtubule is therefore composed
entirely of GTP tubulin subunits, forming what is
known as a GTP caphowever, occasionally
tubulin hydrolyzes its GTP before the next tubulin
has been addedfree ends of protofilaments are
now composed of GDP-tubulin subunitsbalance
in favor of disassemblythe microtubule starts to
shrink. The GDP-containing tubulin molecules
that are freed nowjoin the unpolymerized
tubulin molecules already in the cytosolThe
tubulin molecules joining the pool then exchange
their bound GDP for GTP, thereby becoming
competent again to add to another microtubule

Fig: GTP hydrolysis controls the growth

of microtubules

Microtubules are maintained by a

balance of assembly & disassembly
In a normal cellthe centrosome is shooting out
& retracting the microtubules continuallycan be
prevented from disassemblingif plus end is
permanently stabilized by attachment to another
molecule or cell structureset up a highly
organized system of microtubules. Drugs that
prevent the polymerization or depolymerization
effect on the organization of the cytoskeleton &
behavior of the celle.g. If a cell in mitosis is
exposed to the drug colchicine, the mitotic
spindle rapidly disappears (because tubulin
addition was blocked) unable to partition its
chromosomes.

 The drug taxolbinds to microtubules &

prevents them from losing subunitsthe
microtubules can grow but cannot shrink
also arrests dividing cells in mitosis.
Thus for the spindle to function,
microtubules must be able not only to
assemble but also to disassemble.
Cancer cellsdividing with less control
can sometimes be killed preferentially by
microtubule-stabilizing and microtubuledestabilizing antimitotic drugsthus
drugs including colchicine, taxol,
vincristine, & vinblastine, are used in the

Fig: The selective stabilization of

microtubules can polarize a cell.

Microtubules organize the Interior of the

cell

Cells are able to modify the dynamic instability of their

microtubulese.g. as cells enter mitosismicrotubules
become dynamicswitching b/w growing and shrinking
This enables them to disassemble rapidly and then
reassemble into the mitotic spindlewhen a cell has
differentiatedthe dynamic instability of its microtubules is
often suppressed by proteins that bind to the ends of
microtubules.
Most differentiated animal cells are polarizede.g. nerve
cellsThe cells polarity is a reflection of the polarized
systems of microtubules in its interiorhelp to position
organelles in their required locationthese oriented tracks
the cell is able to send cargoes of materialsmovement
along microtubules is faster than free diffusion. Microtubules
activitydepends on a large variety of accessory proteins
binding themsome stabilize microtubules, link
microtubules to other cell components or are motor proteins

Fig: Microtubules transport

cargo along a nerve cell axon.

Motor Proteins drive Intracellular

transport

Cytoplasmin continual motionMitochondria

& the smaller membrane-enclosed organelles
and vesiclesmove in small, jerky steps
move for a short period, stop, and then start
againsaltatory movementmuch more
sustained and directional than the continual,
small Brownian movements caused by random
thermal motionsBoth microtubules and actin
filaments are involved in itgenerated by
motor proteins, which use the energy derived
from repeated cycles of ATP hydrolysismotor
proteins also attach to other cell components
Dozens of motor proteins.

 The motor proteins that move along cytoplasmic

microtubulese.g. in the axon of a nerve cell
belong to two families: the kinesins (generally
move toward the plus end), while the dyneins
move toward the minus end (toward the
centrosome, inward to the cell body)both are
dimers with two globular ATP-binding heads and a
single tailThe heads interact with microtubules
The tail binds to some cell component, such as
a vesicle or an organelle & thus determines the
type of cargoheads of kinesin & dynein are
enzymes with ATP-hydrolyzing (ATPase) activity
provides the energy for a cycle of conformational
changes in the headenable it to move along the
microtubule by a cycle of binding, release, and
rebinding.

Fig: Motor proteins move along

microtubules using their globular
heads.

Organelles move along

microtubules

Microtubules & motor proteinsimportant part in

positioning membrane-enclosed organellese.g. the
Golgi apparatus is located in the interior of the cell
near the centrosomedepend on microtubules for its
alignment & positioningDyneins, attached to the
Golgi membranespull the Golgi apparatus inward
toward the cell centerregional differences in
internal membranes, on which the successful
function of the cell depends. When cells are treated
with a drug such as colchicineorganelles change
their location dramaticallyGolgi apparatus
fragments into small vesicles, which disperse
throughout the cytoplasmWhen the drug is
removedthe organelle return to its original
positions.

Cilia & Flagella contain stable

microtubules moved by dynein
Stable microtubulesas stiff supportsto construct a
variety of polarized structuresincluding cilia &
flagellato move water over the surface. Ciliahairlike
structures about 0.25 mm in diameter, covered by
plasma membraneextend from the surfacecontains
a core of stable microtubulesthat grow from a basal
body in the cytoplasm; the basal body serves as the
organizing center for the ciliummove fluid or propel
single cells through a fluide.g. protozoa use cilia to
collect food particles, others use them for locomotion.
On the epithelial cells lining the human respiratory
tracthuge numbers of ciliasweep layers of mucus
containing trapped dust particles and dead cells up
toward the throat, to be swallowed & eventually
eliminated from the body.

 The flagellamuch like cilia but are usually much

longerdesigned to move the entire cell
propagate regular waves along their length that
drive the cell through liquid. The microtubules in
cilia & flagella are slightly different from the
cytoplasmic microtubulesA cross section through
a cilium shows nine doublet microtubules arranged
in a ring around a pair of single microtubulesThis
9 + 2 array is characteristic of almost all forms
of eucaryotic cilia & flagella. The movement of a
cilium or a flagellumby the bending of its core
The microtubules are associated with numerous
proteinsproject at regular positions along the
lengthSome serve as cross-links to hold the
bundle of microtubules together; others generate
the force that causes the cilium to bend.

 Ciliary dyneinimportant accessory protein

generates the bending motion of the core
closely resembles cytoplasmic dynein
attached by its tail to one microtubuleheads
interact with an adjacent microtubule to
generate a sliding force b/w the two filaments.
In humanshereditary defects in ciliary dynein
Kartageners syndromeMen with this
disorder are infertile because their sperm are
non-motileaffected have an increased
susceptibility to bronchial infections because
the cilia that line their respiratory tract are
paralyzed and thus unable to clear bacteria &
debris from the lungs.

Fig: Hairlike cilia coat the

surface of many eucaryotic cells.

Fig: Microtubules in a cilium or

flagellum are arranged in a 9 + 2
array.

Fig: The movement of dynein

causes the flagellum to bend.