The cytoskeleton is a network of fibers that organizes structures and activities in the cell The cytoskeleton is a network of fibers extending throughout the cytoplasm It organizes the cells structures and activities, anchoring many organelles It is composed of three types of molecular structures: – Microtubules – Microfilaments – Intermediate filaments

Microtubule Microfilaments 0.25 µm

Roles of the Cytoskeleton: Support, Motility, and Regulation The cytoskeleton helps to support the cell and maintain its shape It interacts with motor proteins to produce motility Inside the cell, vesicles can travel along monorails provided by the cytoskeleton Recent evidence suggests that the cytoskeleton may help regulate biochemical activities

Vesicle ATP Receptor for motor protein Microtubule of cytoskeleton Motor protein (ATP powered) (a) MicrotubuleVesicles (b) 0.25 µm

Components of the Cytoskeleton Three main types of fibers make up the cytoskeleton: – Microtubules are the thickest of the three components of the cytoskeleton – Microfilaments, also called actin filaments, are the thinnest components – Intermediate filaments are fibers with diameters in a middle range

10 µm Column of tubulin dimers Tubulin dimer Actin subunit 25 nm 7 nm Keratin proteins Fibrous subunit (keratins coiled together) 8–12 nm

10 µm Column of tubulin dimers Tubulin dimer 25 nm

Actin subunit 10 µm 7 nm

5 µm Keratin proteins Fibrous subunit (keratins coiled together) 8–12 nm

Microtubules Microtubules are hollow rods about 25 nm in diameter and about 200 nm to 25 microns long Functions of microtubules: – Shaping the cell – Guiding movement of organelles – Separating chromosomes during cell division

Centrosomes and Centrioles In many cells, microtubules grow out from a centrosome near the nucleus The centrosome is a microtubule-organizing center In animal cells, the centrosome has a pair of centrioles, each with nine triplets of microtubules arranged in a ring

Centrosome Microtubule Centrioles 0.25 µm Longitudinal section of one centriole Microtubules Cross section of the other centriole

Cilia and Flagella Microtubules control the beating of cilia and flagella, locomotor appendages of some cells Cilia and flagella differ in their beating patterns

5 µm Direction of swimming (a) Motion of flagella Direction of organisms movement Power strokeRecovery stroke (b) Motion of cilia 15 µm

Cilia and flagella share a common ultrastructure: – A core of microtubules sheathed by the plasma membrane – A basal body that anchors the cilium or flagellum – A motor protein called dynein, which drives the bending movements of a cilium or flagellum

0.1 µm Triplet (c) Cross section of basal body (a)Longitudinal section of cilium 0.5 µm Plasma membrane Basal body Microtubules (b)Cross section of cilium Plasma membrane Outer microtubule doublet Dynein proteins Central microtubule Radial spoke Protein cross- linking outer doublets 0.1 µm

How dynein walking moves flagella and cilia: Dynein arms alternately grab, move, and release the outer microtubules – Protein cross-links limit sliding – Forces exerted by dynein arms cause doublets to curve, bending the cilium or flagellum Copyright © 2008 Pearson Education, Inc., publishing as Pearson Benjamin Cummings

Microtubule doublets Dynein protein ATP (a) Effect of unrestrained dynein movement Cross-linking proteins inside outer doublets Anchorage in cell (b) Effect of cross-linking proteins (c) Wavelike motion

Microtubule doublets Dynein protein (a) Effect of unrestrained dynein movement ATP

Cross-linking proteins inside outer doublets Anchorage in cell ATP (b) Effect of cross-linking proteins (c) Wavelike motion 13 2

Microfilaments (Actin Filaments) Microfilaments are solid rods about 7 nm in diameter, built as a twisted double chain of actin subunits The structural role of microfilaments is to bear tension, resisting pulling forces within the cell They form a 3-D network called the cortex just inside the plasma membrane to help support the cells shape Bundles of microfilaments make up the core of microvilli of intestinal cells

Thinner, shorter and more flexible than microtubules Contains G-actin, and F-actin Actin- most abundant intracellular protein in most Eukaryotic cells Comprises 10% by weight of total cell protein in muscle cells, 1-5% in non-muscle cells Actin polymerization requires K+, Mg 2+, ATP, and Calcium Actin Filaments

G-actin: exists as a globular monomer F-actin: is a helical filamentous polymer of G-actin subunits all oriented in the same direction Microfilaments in a cell are constantly shrinking or growing in length Bundles and meshworks of microfilaments are forming and dissolving continuously G-actin and F-actin

Nucleation - G-actin clumps into short, unstable oligomers, 3-4 subunits long, and acts as a stable seed or nucleus. Elongation phase - The nucleus rapidly increases in length by the addition of actin monomers to both of its ends. Dynamics of Actin Assembly

Steady-State – The ends of actin filaments are in a steady state with monomeric G-actin. – After their incorporation into a filament, subunits slowly hydrolyze ATP and become stable F-actin. Dynamics of Actin Assembly

All subunits in an actin filament point toward the same direction of the filament – Exhibits polarity-actin subunit exposed to the surrounding solution is the (-) end – The cleft that has contact with the neighboring actin subunit that is not exposed is the (+) end – Actin filaments grow faster at the (+) end than the (-) end Dynamics of Actin Assembly

F-actin has structural and functional polarity

Anchorage and movement of membrane proteins- – filaments are distributed in 3-dimensional networks throughout the cell – used as anchors with in specialized cell junctions Actin Filaments participate in a variety of cell functions:

Formation of the structural core of microvilli – On epithelial cells, help maintain shape of the cell surface Actin Filaments participate in a variety of cell functions:

Locomotion of the cells – Achieved by the force exerted by actin filaments by polymerization at their growing ends – Used in many migrating cells, particularly on transformed cells of invasive tumors – Cells extend processes from their surface by pushing the plasma membrane ahead of the growing actin filaments Actin Filaments participate in a variety of cell functions:

Essential in cytoplasmic streaming

Functions and structure of intermediate filaments distinguish them from other cytoskeletal fibers

Microvillus Plasma membrane Microfilaments (actin filaments) Intermediate filaments 0.25 µm

Microfilaments that function in cellular motility contain the protein myosin in addition to actin In muscle cells, thousands of actin filaments are arranged parallel to one another Thicker filaments composed of myosin interdigitate with the thinner actin fibers

Muscle cell Actin filament Myosin filament Myosin arm (a) Myosin motors in muscle cell contraction Cortex (outer cytoplasm): gel with actin network Inner cytoplasm: sol with actin subunits Extending pseudopodium (b) Amoeboid movement Nonmoving cortical cytoplasm (gel) Chloroplast Streaming cytoplasm (sol) Vacuole Cell wall Parallel actin filaments (c) Cytoplasmic streaming in plant cells

Muscle cell Actin filament Myosin filament Myosin arm (a) Myosin motors in muscle cell contraction

Cortex (outer cytoplasm): gel with actin network Inner cytoplasm: sol with actin subunits Extending pseudopodium (b) Amoeboid movement Nonmoving cortical cytoplasm (gel) Chloroplast Cell wall Streaming cytoplasm (sol) Parallel actin filaments (c) Cytoplasmic streaming in plant cells Vacuole

Localized contraction brought about by actin and myosin also drives amoeboid movement Pseudopodia (cellular extensions) extend and contract through the reversible assembly and contraction of actin subunits into microfilaments

Cytoplasmic streaming is a circular flow of cytoplasm within cells This streaming speeds distribution of materials within the cell In plant cells, actin-myosin interactions and sol- gel transformations drive cytoplasmic streaming