CYTOSKELETON

Norhaifah S. Tacoranga
 CYTOSKELETON
A system of filaments or fibres that is
present in the cytoplasm of eukaryotic
cells, prokaryotic cells and archeans.
First described in erythrocytes, where it
maintains the cellular biconcave shape
and gives the cell its properties of
elasticity and flexibility. It is a polymer
network, composed of three distinct
biopolymer types: the actin filaments
(microfilaments), microtubules,
intermediate filaments.
 Functions of the Cytoskeleton
• It provides shape and support to the cell.
• It helps in the formation of vacuoles.
• It holds different cell organelles in place.
• It assists in cell signalling.
• It supports intracellular movements like the
migration of cell organelles, transportation of
vesicles in and out of the cell, etc.
 Structure of the Cytoskeleton
• Actin Filaments (Microfilaments)
• Microtubules
• Intermediate Filaments
• Motor Proteins
CYTOSKELETAL
FILAMENTS
 ACTIN FILAMENTS
Actin Filaments (AF) have varying sizes according
to different resources. There were microfilaments
described to have a 3-6 nm in diameter and
microfilaments described to have 7- to 8-nm diameter.
They are built from dimer pairs of globular actin
monomers, and are functionally polar in nature. They
have two distinct ends: a fast and a slow growing end
(called the plus end and minus end respectively).
Actin Filament assembly
starts from activation
and ends with the
deactivation of G-actins.
Actin Filament formation
maintenance and the proteins
cycles involved in the process (e.g.
Cofilin Cycle, Profilin Cycle, and
Thymosin Beta4 Cycle).
Actin Filaments
undergo
polymerization
or
depolymerization
depending on the
monomeric actin
concentration it
encounters.
Actin Filaments can grow asymmetrically. And when the actin monomer concentration lies
between the two values, only the plus end grows while the minus end shrinks. This process,
where the length of the filament stays roughly constant and the polymerized monomers
within the AF transfer momentum forward due to asymmetric plus end polymerization, is
known as treadmilling; it is a critical aspect of how polymerizing AFs can generate force.
 MICROTUBULES
Microtubules (MTs) are the stiffest of the biopolymers, with
Lp ranging from 100 to 5000 µm depending on the filament
length. MTs are rod-like polymers, with an outer diameter of ~25
nm. It is coprised of Tubulin protein subunits that assemble into
protofilaments, and typically 13 of these protofilaments then
align to form a hollow tube, imbuing MTs with their incredible
rigidity. MTs exhibit similar dynamics to those of actin: they are
functionally polar, treadmill, and can impart a force through
polymerization. They are constantly assembling and
disassembling.
 INTERMEDIATE FILAMENTS
Intermediate filaments (IFs) are much more flexible than
AFs and MTs (Lp~0.3-1.0 µm). They range in diameter from 8
to12 nm, between that of AFs and MTs. There are different
classes of IFs such as vimetin, desmin, keratin, lamin and
neurofilaments, with different cell types having different IFs.
Unlike AFs or MTs, IFs are not polarized, do not treadmill, do
not generally depolymerize under physiological conditions
once polymerized, and are therefore considered to be more
static in nature than AFs and MTs.
Intermediate Filament
formation starts from
the crosslinking of two
monomers that will
form a single unit.
MOTOR PROTEINS
 ACTIN-BASED MOTORS
Myosins are composed of three common domains 5: (1) the N-
terminal motor domains that interact with actin and hydrolyze ATP to
generate force, (2) the neck domains or lever arms that transduce and
amplify the force generated by motor domain, and (3) the tail domains
offer specific binding sites for different cargos. The motor domain of
myosin has an actin-binding site and a nucleotide-binding site, which are
mutually exclusive. Following the motor domain is lever armThe length
of lever arm determines the step size along actin. The most essential
difference among myosin motor proteins lies in their C-terminal globular
tail domain (GTD). This domain recognizes various cargos through
direct interactions or via adaptor proteins.
 MICROTUBULE-BASED MOTORS
Kinesins (also referred to as KIFs) propel cargos along the microtubule.
All kinesins contain a highly conserved globular motor domain comprising a
microtubule-binding site and an ATP-binding site as well as a diverse tail
domain, which is responsible for cargo recognition and binding via the
adaptor or scaffolding proteins. Unlike myosin whose motor domain is
always located in N-terminus, the motor domain of kinesins can be found in
their N-terminal (N-type), middle (M-type), or C-terminal (C-type) region.
On microtubule, N-type kinesin moves to the plus end of microtubule, C-
type kinesin moves to minus end of microtubule and M-type kinesin can
destabilize microtubule tracks. The energy supplying for kinesin processive
motility comes from hydrolyzing ATP in the motor domain.
MICROTUBULE-BASED MOTORS
 MICROTUBULE-BASED MOTORS
Another microtubule-based motor is cytoplasmic dynein. Compared
to myosin and kinesin, dynein is larger in size with more complicated
composition and its working mechanisms are not well understood. The
motor domain of dynein is located in C-terminal region, while tail
domain is situated in the N-terminal region. Microtubule-binding
domain, coupled with ATP-hydrolysis cycle, moves the protein towards
minus-end microtubule cytoskeleton. The dynein tail domain is
responsible for dynein homodimerization and serves as binding sites
with other subunits of dynein. These dynein subunits can interact with
adaptor proteins to form bridges. With these bridges, dynein tail
domain can thus carry the target cargos.
CELL MOTILITY
 CELL MOVEMENT
A cell begins to move in
response to an external signal in its
surrounding environment. This can
be a physical, chemical, diffusible
or non-diffusible signal that is
detected by receptor proteins
located on the cell membrane, and
transmitted by them via signaling
cascades to the cell interior.
 CELL MOVEMENT
The process can be divided into three
stages in most cells. First, a cell propels the
membrane forward by orienting and
reorganizing (growing) the actin network at
its leading edge. Second, it adheres to the
substrate at the leading edge and
deadheres (releases) at the cell body and
rear of the cell. Finally, contractile forces,
generated largely by the action of the acto-
myosin network, pull the cell forward.
 CELL MOVEMENT
After sensing the signal, the cell starts
moving in response to the signal by
polymerizing actin. As the extending edge
moves forward, the cell constantly monitors
the signal direction and tailors its direction
of motion accordingly. Soon after the
leading edge begins to protrude, adhesion
molecules gathered in the extending region
help attach the leading edge to the substrate.

Note: Chemoreactant and Chemorepellant

CELL JUNCTIONS
CELL JUNCTION CLASSIFICATION
 OCCLUDING JUNCTIONS
Tight Junctions are composed
of a branching network of sealing
strands that completely encircles
the apical end of each cell in the
epithelial sheet. Each tight
junction sealing strand is
composed of a long row of
transmembrane adhesion proteins
embedded in each of the two
interacting plasma membranes.
 ANCHORING JUNCTIONS
Anchoring junctions are widely distributed in animal
tissues and are most abundant in tissues that are
subjected to severe mechanical stress. They are composed
of two main classes of proteins: Intracellular anchor
proteins and Transmembrane adhesion proteins In
addition to anchor proteins and adhesion proteins, many
anchoring junctions contain intracellular signaling proteins
that enable the junctions to signal to the cell interior. They
can be classified into integrins and cadherins.
 ANCHORING JUNCTIONS
Anchoring junctions have to functional
forms: Adherens junctions and desmosomes and
Focal adhesions and hemidesmosomes.
Adherens junctions and desmosomes and
connect cells together and are formed by
cadherins, while focal adhesions and
hemidesmosomes connect cells to the
extracellular matrix and are formed by integrins.
Adherens junctions
and focal adhesions
are connecting sites
for bundles of actin
filaments, whereas
desmosomes and
hemidesmosomes
are connecting sites
for intermediate
filaments.
 GAP JUNCTIONS
A gap-junction connexon is made up of six
transmembrane connexin subunits. Connexins are
four-pass transmembrane proteins, six of which
assemble to form a channel, a connexon. When the
connexons in the plasma membranes of two cells in
contact are aligned, they form a continuous aqueous
channel that connects the two cell interiors. The
connexons hold the interacting plasma membranes
at a fixed distance apart—hence the gap.
The channels that
connexons form allow
inorganic ions and
other small water-
soluble molecules to
pass directly from the
cytoplasm of one cell
to the cytoplasm of
the other, thereby
coupling the cells
both electrically and
metabolically.
THAT IS ALL, THANK YOU.