1. 02.16.11 Lecture 12 - The actin cytoskeleton

2. Actin filaments allow cells to adopt different shapes and perform different functions VilliContractile bundles Sheet-like & Finger-like protrusions Contractile ring

3. Actin filaments are thin and flexible 7 nm in diameter Less rigid than microtubules Plus end - fast growing Minus end - slow growing Monomers polymerize into a helical chain

4. Actin and microtubules polymerize using similar mechanisms Monomeric actin binds to ATP Upon polymerization, actin ATPase activity cleaves ATP to ADP ATP hydrolysis acts as a molecular “clock” Older actin filaments with ADP are unstable and disassemble

5. Actin architecture and function is governed by actin-binding proteins

6. Example: actin in microvilli

7. Example: actin in the cell cortex

8. Actin polymerization can produce “pushing” forces Polymerization at the front of a cell pushes the leading edge forward Phagocytosis - formation of pseudopods Intracellular movement and cell-to-cell spreading of pathogens

9. During cell migration, actin polymerization pushes the leading edge forward

10. Actin polymerization drives protrusion of the cell membrane Lamellipodia Filopodia

11. 11 Lamellipodia are composed of branched networks of short filaments

12. Model for actin polymerization at membranes in lamellipodia

13. Filopodia are composed of long, unbranched actin filaments

14. Actin polymerization powers engulfment during phagocytosis

15. Movement of Listeria monocytogenes Pathogenic bacterium that colonizes the epithelial cells lining the gut Found in contaminated dairy products Infection can be lethal to newborns and immunocompromised individuals

16. Listeria move on an actin-based “comet- tail”

17. Myosins are actin-based motor proteins Myosins convert ATP hydrolysis into movement along actin filaments Many different classes of myosins (>30 in humans) Some myosins move cargoes, other myosins slide actin (as in muscles) Actin & ATP binding sites in N-terminal head domain

18. Myosins “walk” along actin filaments

19. Myosin I can carry organelles or slide actin filaments along the membrane

20. Myosin II slides actin filaments to produce contractile forces

21. Myosin-based contraction drives cytokinesis

22. Skeletal muscle cells are packed with myofibrils, each of which contains repeating chains of sarcomeres

23. Sarcomeres are contractile units of actin and myosin II

24. In muscle cells, myosin II is a filament of many motors

25. Muscle contraction is driven by myosin II

26. The myosin cycle in muscle

27. Contraction is activated by calcium release from the sarcoplasmic reticulum

28. Calcium release channels are opened by a voltage-sensitive transmembrane protein in the T-tubule

29. Contraction is regulated by a Ca+2-mediated change in the conformation of troponin

30. Muscle contraction