Review the Concepts

1. Microtubules are polar filaments; that is, one end is different from the other. What is the basis for this polarity, how is polarity related to microtubule organization within the cell, and how is polarity related to the intracellular movements powered by microtubule-dependent motors?

2. Microtubules both in vitro and in vivo undergo dynamic instability, and this type of assembly is thought to be intrinsic to the microtubule. What is the current model that accounts for dynamic instability?

3. In cells, microtubule assembly depends on other proteins as well as tubulin concentration and temperature. What types of proteins influence microtubule assembly in vivo, and how does each type affect assembly?

4. Microtubules within a cell appear to be arranged in specific arrays. What cellular structure is responsible for determining the arrangement of microtubules within a cell? How many of these structures are found in a typical cell? Describe how such structures serve to nucleate microtubule assembly.

5. Many drugs that inhibit mitosis bind specifically to tubulin, microtubules, or both. What diseases are such drugs used to treat? Functionally speaking, these drugs can be divided into two groups based on their effect on microtubule assembly. What are the two mechanisms by which such drugs alter microtubule structure?

6. Kinesin-1 was the first member of the kinesin superfamily to be identified and therefore is perhaps the best-characterized superfamily member. What fundamental property of kinesin was used to purify it?

7. Certain cellular components appear to move bidirectionally on microtubules. Describe how this is possible given that microtubule orientation is fixed by the MTOC.

8. The movement of kinesin motor proteins involves both the motor domain and the linker domain. Describe the role of each domain in kinesin movement, direction of movement, or both. Could kinesin-1 with one inactive head efficiently move a vesicle along a microtubule?

9. What features of the dynactin complex enable cytoplasmic dynein to transport cargo toward the microtubule (−) end? What effect could inhibition of dynactin interaction with the +TIP EB1 have on spindle orientation?

10. Cell swimming depends on appendages containing microtubules. What is the underlying structure of these appendages, and how do these structures generate the force required to produce swimming?

11. What effect would dynein inactivation have on kinesin-2-dependent IFT?

870

12. The mitotic spindle is often described as a microtubule-based cellular machine. The microtubules that constitute the mitotic spindle can be classified into three distinct types. What are the three types of spindle microtubules, and what is the function of each?

13. Mitotic spindle function relies heavily on microtubule motors. For each of the following motor proteins, predict the effect on spindle formation, function, or both of adding a drug that specifically inhibits only that motor: kinesin-5, kinesin-13, and kinesin-4.

14. The poleward movement of kinetochores, and hence chromatids, during anaphase A requires that kinetochores maintain a hold on the shortening microtubules. How does a kinetochore hold onto shortening microtubules?

15. Anaphase B involves the separation of spindle poles. What forces have been proposed to drive this separation? What underlying molecular mechanisms are thought to provide these forces?

16. Cytokinesis, the process of cytoplasmic division, occurs shortly after the separated sister chromatids have neared the opposite spindle poles. How is the plane of cytokinesis determined? What are the respective roles of microtubules and actin filaments in cytokinesis?

17. The best strategy for treating a specific type of human tumor can depend on identifying the type of cell that became cancerous to give rise to the tumor. For some tumors that have colonized a distant location (metastasized), identifying the parental cell type can be difficult. Because the type of IF protein expressed is cell-type-specific, using monoclonal antibodies that react with only one type of IF protein can help in this identification. What IF proteins would you produce monoclonal antibodies against to identify (a) a sarcoma of muscle cell origin, (b) an epithelial cell carcinoma, and (c) an astrocytoma (glial cell tumor)?

18. Explain why there are no known motors that use intermediate filaments as tracks.

19. Growth cones are highly mobile regions of developing neurons. What prevents the growth cone from moving or collapsing back into the main cell body, as often occurs with lamellipodia?