m The cell interior is highly ordered as a result of the

cytoskeleton.
m The cytoskeleton is a filamentous network of protiens
that pervades the cytoplasm
m The protein filaments consist of protein monomers that
self-assemble into the final structure, and can
dissociate to destroy that structure.
m In general, these filaments are highly dynamic - they
are not permanent or static structures.
m Providing structural support to the cell, the
cytoskeleton also functions in cell motility and
regulation.
m á-12 nm in diameter (Ú
Ú

between microtubules and actin)
m The most stable and least dynamic
of the filaments
m Filaments are not polarized
m Thought to serve a strengthening
and stabilizing role in cells
m Central rods (alpha helix, coiled coil
dimer)
m Dimer± to tetramer (antiparallel
structure)
m Tetramer connected longitudinally
(protofilaments)
m Eight protofilaments make one
filament

The monomer is not globular but fibre.

m more permanent fixtures
of the cytoskeleton than
are the other two classes
m reinforce cell shape
m and fix organelle location.
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m the thickest fibers, are hollow rods about 25 microns
in diameter.

m microtubules are dynamic structures that undergo

continual assembly and disassembly within the cell.

m They function both to determine cell shape and in a

variety of cell movements, including some forms of
cell locomotion, the intracellular transport of
organelles, and the separation of chromosomes
during mitosis.
m Tubes made of Į- and ȕutubulin
m Exist Ú Ú as heterodimers
m Form protofilaments, which
circularize in groups of 13
(usually)
m One end is all alpha (+ end),
the other all beta (- end)« that
is, they are polarized
m ` 
, growth occurs
preferentially at the + end
m echanically analogous to rigid
rods
m polarity is an important
consideration in determining
the direction of movement
along microtubules
m icrotubules are nucleated by microtubule
organizing centers (TOCs)
m The centrosome is an example of a TOC
m TOCs contain microtubule structures
that form stable (-) ends for growing microtubules
m Contain Ȗ-tubulin which may stabilize
these nascent microtubules
The centrosomes of most animal cells contain a pair of centrioles, oriented
perpendicular to each other, surrounded by amorphous pericentriolar material.
The centrioles are cylindrical structures consisting of nine triplets of
microtubules
$(!{%&){ *{($+{!$ ( ,$% !)-)&

The minus ends of microtubules are anchored in the centrosome. In

interphase cells, the centrosome is located near the nucleus and
microtubules extend outward to the cell periphery. During mitosis,
duplicated centrosomes separate and microtubules reorganize to form the
mitotic spindle. Chromosome movement is mediated by motor proteins
associated with the spindle microtubules
m Tubulin dimers bind GTP
m Tubulin-GTP adds to the (+) end of the
filament
m Once in the microtubule, the GTP is almost
immediately hydrolyzed, leaving a *!"
on the (+) end.
m This GTP hydrolysis weakens the binding
affinity of tubulin for adjacent molecules,
thereby favoring depolymerization and
resulting in the dynamic behavior of
microtubules.
m Whether a microtubule grows or shrinks is
determined by the rate of tubulin addition
relative to the rate of GTP hydrolysis. As long
as new GTP-bound tubulin molecules are
added more rapidly than GTP is hydrolyzed,
the microtubule retains a GTP cap at its plus
end and microtubule growth continues.
However, if the rate of polymerization slows,
the GTP bound to tubulin at the plus end of
the microtubule will be hydrolyzed to GDP. If
this occurs, the GDP-bound tubulin will
dissociate, resulting in rapid depolymerization
and shrinkage of the microtubule.
m any chemotherapeutic drugs are antimitotics (shut
down mitosis)
D vinblastine causes tubulin aggregation
D nocodazole caps microtubules so that they can¶t grow
D taxol stabilizes microtubules, blocking complete cell
division
m colchicine
D from the Autumn Crocus (a lavender)
D causes disassembly of microtubules
m aometimes used to enhance the potency of
marijuana
D Causes some chromosomes to not always be evenly
divided between daughter cells
D Causes some cells to become polyploid
D induces overexpression of genes.
m the thinnest class of the cytoskeletal
fibers 7 um.
m are solid rods of the globular protein

.
m designed to resist tension
m form a three-dimensional network just
inside the plasma membrane where they
form a network that provides mechanical
support, determines cell shape, and
allows movement of the cell surface,
thereby enabling cells to migrate, engulf
particles, and divide.
m Over 5% of the intracellular protein
m The most ubiquitous of all intracellular proteins
m Functions
D Cell motility and muscle
contraction
D Cell shape (like microvilli)
D Vesicle transport near cell
periphery
m Actin monomers stack to form a double-helical
filament, 7 nm in width
m While microtubules are º
ases, actin filaments
are 
ases
m The + end grows faster than the ± end

m Once in the filament, the rate of ATP hydrolysis

increases

Actin monomers 
 u

Actin-ATP

 u
 Actin-ADP
Actin filaments in bundles are crosslinked into parallel arrays by small
proteins that align the filaments closely with one another. In contrast,
networks are formed by large flexible proteins that crosslink orthogonal
filaments.
m In muscle cells, thousands of actin filaments are
arranged parallel to one another.
m Thicker filaments, composed of a motor protein, 

,
interdigitate with the thinner actin fibers
m In other cells, these actin-myosin aggregates
are less organized but still cause localized
contraction
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m In plant cells (and others), actin-myosin
interactions and sol-gel transformations drive


 
.
m Phallotoxin (phalloidin)
D an actin filament stabilizer
D the poison in some mushroom genera
D It kills by 
 ÚÚÚ  actin
filaments (inhibiting disassembly)
D Immediate cause of death is
liver failure
m Cytochalasin
D an actin filament de-stabilizer
D also derived from mushrooms

 

 
m With few exceptions, plus-end
directed
m yosins have heads that serve the
motor-function, and tails that either:
D Form filaments, as in muscle
D Bind cargo, like kinesins and dynein
m Two major kinds:
D üinesins (most are plus-
end directed)
D Dyneins (minus-end
directed)
m Both have motor
domains (heads) and
cargo-binding domains
(tails)
m Dyneins are of two kinds
D Cytoplasmic ± cargo
transport
D Axonemal ± ciliary and
flagellar motion
m Otherwise you would rely on diffusion
m The distance traveled by diffusion is
linearly
related to time!
Thymosin is a monomer buffer
Profilin is a shuttle, and promotes nucleotide
exchange
m Jamellipodia and pseudopodia are pushed forward
by actin polymerization and other forces
m How does a cancer cell or leukocyte crawl ³in the
right direction,´ like
 a cytokine?
R

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m In many cells,  


in the EC connect to

 

, intrinsic membrane proteins.
m Plant cells are perforated with 

,
channels allowing cysotol to pass between cells.
m In .


, membranes of adjacent
cells are fused, forming continuous belts
around cells.
D This prevents leakage of extracellular fluid.
m *.


(or communicating
junctions) provide cytoplasmic
channels between adjacent cells.
m In many cells, microtubules grow out from a


 near the nucleus.
m `






  










 



 

 







m 







 
 
m icrotubules are the central structural support
m Cilia usually occur in large numbers on the cell
surface.
m There are usually just one or a few flagella per
cell% and   are microtubule-based
projections of the plasma membrane that are
responsible for movement of a variety of
eukaryotic cells.
m A flagellum has an undulatory movement
m Cilia move more like oars with alternating power
and recovery strokes.
m have the same ultrastructure.