MOLECULAR CELL BIOLOGY

BS101

12-13
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Lecture 8 The cytoskeleton and cytosol, microfilaments and microtubules

LEARNING OBJECTIVES

Lecture 8

List and describe the molecules that compose the cellular cytoskeleton.
Describe the main functions of the cytoskeleton.

Original views on cell structure

In the early days of light microscopy, scientists thought that organelles floated freely in the cytoplasm of a cell Eventually, improvements in both light and electron microscopy revealed that the cell contained a CYTOSKELETON

Cytoskeleton
Green= microtubules Red= actin filaments

Blue= DNA in nucleus

This shows that the cytoskeleton extends throughout the cell It acts like a dome tent, stabilised by opposing forces But it also very dynamic, and can be dismantled and reassembled quickly, to change the shape of the cell

Figure 6.20 Reece et al (2011)

The cytoskeleton is a network of fibres that organizes structures and activities in the cell
The cytoskeleton is a network of fibres extending throughout the cytoplasm, providing mechanical support and to maintain the cell shape It organizes the cells structures and activities, anchoring many organelles It is composed of three types of molecular structures: Microtubules Microfilaments Intermediate filaments
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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

ATP

Vesicle Receptor for motor protein

This shows how vesicles can be carried around the cell on monorails of microtubules Here, motor proteins can attach themselves to a vesicle, then also to a microtubule, and using ATP, this motor protein can walk the vesicle along the rail, to its destination with the cell In this figure of a squid giant axon, two vesicles containing neurotransmitters migrate toward the tip of the axon, by the same mechanism as above

Motor protein (ATP powered) (a) Microtubule Vesicles

Microtubule of cytoskeleton

0.25 m

(b)

Reece et al (2011) Fig. 6-21

Kinesin walking along a microtubule http://www.youtube.com/watch?v=YA va4g3Pk6k

Movement of vesicles within the cell

Vesicles of proteins that leave the ER and move to the Golgi (endomembrane system lecture 6) are transported along cytoskeletal tracks The cytoskeleton also manipulates the plasma membrane, making it bend inward to form food vacuoles or phagocytic vesicles

Components of the Cytoskeleton

Three main types of fibres 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 fibres with diameters in a middle range

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Campbell and Reece (2011)

Table 6.1 Structure and function of the cytoskeleton

10 m

10 m

10 m

Microfilaments Intermediate filaments Microtubules

Column of tubulin dimers Actin subunit

25 nm 7 nm

Keratin proteins Fibrous subunit (keratins coiled together)

812 nm

Tubulin dimer

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10 m

Dimer of and tubulin

e.g.Flagella/cilia Column of tubulin dimers

e.g. mitotic spindle e.g. movement of secretory vesicles from Golgi to membrane surface

25 nm

Campbell and Reece (2011) Table 6-1a

Tubulin dimer

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Microtubule functions
25nm diameter Organising role

Grow out from a centrosome

Provide a system of tracks, along which other organelles and vesicles are moved

Also responsible for anchoring organelles within the cell

Form the mitotic spindle during cell division Also present in cilia and flagella for movement (e.g. sperm)
Figure 17-8 Essential Cell Biology ( Garland Science 2010)

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Centrosomes and Centrioles

In many cells, microtubules grow out from a centrosome near the nucleus The centrosome is a microtubule-organizing centre 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 m

Most animal cells have a centrosome- a region near the nucleus where the cells microtubules are initiated Within the centrosome is a pair of centrioles: each 0.25um in diameter The two centrioles are at right angles to each other, and each is made up of 9 sets of three microtubules

Longitudinal section of Microtubules one centriole

Cross section of the other centriole

Reece et al (2011) Fig. 6-22

Campbell and Reece (2011) Figure 12.8 The mitotic spindle at metaphase

-condensed chromatin -mitotic spindle is made of tubulin (microtubules) -mitotic spindle radiates out from the centrosome -chromosomes bind to spindle via kinetochores - the kinetochore is bound to centromeric DNA
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Cilia and Flagella

Microtubules control the beating of cilia and flagella, locomotor appendages of some cells Cilia and flagella differ in their beating patterns Many unicellular organisms are propelled through water by cilia or flagella
06_23aC hlamy domonas_SV .mpg

Chlamydomonas (green unicellular algae) showing moving flagella

Cilia and flagella

Cilia can be found on the surface of a cell, and can move liquids along E.g. ciliated lining of trachea can move mucus and dust particles out of lungs Cilia in oviducts of womens reproductive tract can move egg towards uterus Cells can have many cilia, but only 1 or 2 flagella The beating patterns of cilia and flagella is different

Direction of swimming

The motion of flagella is undulating, like a snake, driving the cell forward in the direction it is facingfor example, this sperm cell

(a) Motion of flagella 5 m

Direction of organisms movement

Power stroke

Recovery stroke

(b) Motion of cilia

Reece et al (2011) Fig. 6-23

15 m

The motion of cilia is different however- they have a back and forth motion, which moves the cell sideways- its like one side on a boat oar The image shows a Colpidium- a freshwater protozoa

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 Arranged in a 9+2 pattern
Animation: Cilia and flagella

In Mastering Biology Study Area, Chapter 6, Activity: Cilia and Flagella

0.1 m

Outer microtubule doublet Dynein proteins Central microtubule Radial spoke

Plasma membrane

Microtubules

(b) Cross section of cilium: Shows a cross section through

the middle of a cilium, showing the 9+2 arrangement of microtubules- the outer ring of doublets are held together by motor proteins called DYENINS (red) The central doublet is held to the outer ring by other cross linking proteins (blue)

Protein crosslinking outer doublets

Plasma membrane Basal body

0.5 m

0.1 m Triplet

(a) Longitudinal section of cilium: showing microtubules running the length of the cilium

Reece et al (2011) Fig. 6-24

(c) Cross section of basal body:

The bottom part of the cilia- each doublet joins up to another microtubule to form a triplet. The central doublet has gone and the structure is held together by the cross linking proteins (blue)

10 m

e.g. interaction with myosin in sarcomere Actin subunit

Campbell and Reece (2011) Table 6-1b

7 nm
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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

Microfilament/Actin filament functions

7nm diameter Actin filaments are thin and flexible, provide shape to the cell A shows microvilli B shows contractile bundles in cytoplasm C shows fingerlike protrusions from the leading edge of a moving cell D shows contractile ring/cleavage furrow during cytokinesis
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Figure 17-28 Essential Cell Biology ( Garland Science 2010)

Microvillus

This image shows the structural role of microfilaments This shows microvilli in the intestine, and inside the microvilli are microfilaments (actin filaments)

Plasma membrane

Microfilaments (actin filaments)

Intermediate filaments

Reece et al (2011) Fig. 6-26

0.25 m

Microfilament/Actin filament functions

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 fibres

Muscle cell
Actin filament (yellow)

In muscle cells during contraction, myosin arms walk over the actin filaments- this causes shortening of the gaps between the actin filaments A muscle contraction involves shortening of many muscle cells at the same time

Myosin filament (purple) Myosin arm (a) Myosin motors in muscle cell contraction

Reece et al (2011) Fig, 6-27a

Cytoplasmic streaming
Is a circular flow of cytoplasm within cells This streaming speeds distribution of materials within the cell Cytoplasmic streaming is a way of stirring up the contents of the cytoplasm In plant cells, actin-myosin interactions drive cytoplasmic streaming

Video: Cytoplasmic Streaming

5 m

e.g. skin, hair, nails

Keratin proteins :structural filaments that help to keep the shape of the nucleus, and are depolymerised during mitosis, when the nuclear envelope disintegrates Campbell and Reece (2011) Table 6-1c
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Fibrous subunit (keratins coiled together) 812 nm

Intermediate Filaments
Intermediate filaments range in diameter from 812 nanometres, larger than microfilaments but smaller than microtubules They support cell shape and fix organelles in place They also bear tension in the cell- and nucleus sits in a cage of intermediate filaments that hold it in centre of cell Intermediate filaments are more permanent cytoskeleton fixtures than the other two classes Eg keratin framework still remains in place, even after skin cells have died

Intermediate filament functions

10nm diameter Intermediate filaments have great tensile strength Allow cells to withstand mechanical stress when stretched They span the entire cell, strengthening it They are the toughest and most durable of the 3 filament types

Figure 17-4 Essential Cell Biology ( Garland Science 2010)

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Microtubules can be targets for anti-cancer drugs

(Yew tree) (Crocuses)

(Madagascar periwinkle)

Cancer cells divide rapidly and with less control Inhibition of the mitotic spindle can prevent cell division, and cause death of these cells These drugs block microtubule polymerisation- anti-mitotic agents

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Table 17-1 Essential Cell Biology ( Garland Science 2010)

LEARNING OBJECTIVES

Lecture 8

List and describe the molecules that compose the cellular cytoskeleton. Describe the main functions of the cytoskeleton.

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Homework: Go to Mastering Biology BS101 account online Complete assignment 4 Cytoskeleton and cytosol

Opens: Tues 23rd Oct, 5pm Closes: Tues 6th Nov, 5pm

Reading: Campbell Biology 9th Ed. Reece et al (2011). Ch 6 pp.158-166 Homework: watch http://www.bbc.co.uk/iplayer/episode/b01nln7d/Secret_Universe_The_ Hidden_Life_of_the_Cell/ Only available until 8pm, Sunday 28th Oct
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