Abstract
Microtubule-associated proteins (MAPs) can promote microtubule assemblyin vitro. One of these MAPs (MAP2) consists of a short promoter domain which binds to the microtubule and promotes assembly and a long projection domain which projects out from the microtubule and may interact wth other cytoskeletal elements. We have previously shown that MAP2 and another MAP, tau, differ in their interactions with tubulin in that tau, but not MAP2, promotes extensive aggregation of tubulin into spiral clusters in the presence of vinblastine and that microtubules formed with MAP2 are more resistant than those formed with tau to the antimitotic drug maytansine [Luduena, R. F.,et al. (1984),J. Biol. Chem. 259, 12890–12898; Fellous, A.,et al. (1985),Cancer Res. 45, 5004–5010]. Here we have used chymotryptic digestion to remove the projection domain of MAP2 and examined the interaction of the digested MAP2 (ctMAP2) with tubulin in the presence of vinblastine and maytansine. We have found that ctMAP2 behaves very much like tau, but not like undigested MAP2, in the presence of vinblastine, in that ctMAP2 causes tubulin to polymerize into large clusters of spirals. In contrast, microtubule assembly in the presence of ctMAP2 is much more resistant to maytansine inhibition than is assembly in the presence of tau or undigested MAP2. Our results suggest that the projection domain of MAP2 may play a role in the interaction of tubulin with MAP2 during microtubule assembly.
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Abbreviations
- MAPs:
-
microtubule-associated proteins
- ctMAP2:
-
MAP2 digested withα-chymotrypsin
- nMAP2:
-
untreated MAP2
- PMSF:
-
phenylmethylsulfonyl fluoride
- GMPCPP:
-
guanosine-5′-(α,β-methylene)triphosphate
References
Bhattacharyya, B., and Wolff, J. (1977).FEBS Lett. 75, 159–162.
Bloom, G. S., and Vallee, R. B. (1983).J. Cell Biol. 96, 1523–1531.
Chen, J., Kanai, Y., Cowan, N. J., and Hirokawa, N. (1992).Nature 360, 674–677.
Doll, T., Papandrikopoulou, A., and Matus, A. (1990).Nucleic Acids Res. 18, 361.
Ennulat, D. J., Liem, R. K. H., Hashim, G. A., and Shelanski, M. L. (1989).J. Biol. Chem. 264, 5327–5330.
Fellous, A., Francon, J., Lennon, A.-M. and Nunez, J. (1977).Eur. J. Biochem. 78, 167–174.
Fellous, A., Luduena, R. F., Prasad, V., Jordan, M. A., Anderson, W., Ohayon, R., and Smith, P. T. (1985).Cancer Res. 45, 5004–5010.
Friedrich, P., and Aszodi, A. (1991).FEBS Lett. 295, 5–9.
Garner, C. C., and Matus, A. (1988).J. Cell Biol. 106, 779–784.
Goedert, M., and Jakes, R. (1990).EMBO J. 9, 4225–4230.
Goedert, M., Spillantini, M. G., Potier, M. C., Ulrich, J., and Crowther, R. A. (1989a).EMBO J. 8, 393–399.
Goedert, M., Spilllantini, M. G., Jakes, R., Rutherford, D., and Crowther, R. A. (1989b).Neuron 3, 519–526.
Goedert, M., Jakes, R., Spillantini, M. G., and Crowther, R. A. (1994). InMicrotubules (Hyams, J. S., and Lloyd, C. W., eds.), Wiley-Liss, New York, pp. 183–200.
Goode, B., and Feinstein, S. (1994).J. Cell Biol. (in press).
Gottlieb, R. A. and Murphy, D. B. (1985).J. Cell Biol. 101, 1782–1789.
Griffith, L. M., and Pollard, T. D. (1982).J. Biol. Chem. 257, 9143–9151.
Himmler, A., Drechsel, D., Kirschner, M. W., and Martin, D. W. (1989).Mol. Cell. Biol. 9, 1381–1388.
Hirokawa, N., Hisanaga, S.-I., and Shiomura, Y. (1988).J. Neurosci. 8, 2769–2779.
Joly, J. C., Flynn, G., and Purich, D. L. (1989).J. Cell Biol. 109, 2289–2294.
Kosik, K. S., Orrechio, L. D., Bakalis, S., and Neve, R. L. (1989).Neuron 2, 1389–1397.
Kotani, S., Kawai, G., Yokoyama, S., and Murofushi, H. (1990).Biochemistry 29, 10049–10054.
Kreis, T., and Vale, R., eds. (1993).Guidebook to the Cytoskeletal and Motor Proteins, Oxford University Press, Oxford, pp. 99–134.
Laemmli, U. K. (1970).Nature 227, 680–585.
Lee, G., Cowan, N., and Kirschner, M. (1988).Science 239, 285–288.
Lewis, S. A., Wang, D., and Cowan, N. J. (1988).Science 242, 936–939.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951).J. Biol. Chem. 193, 265–275.
Luduena, R. F., Fellous, A., Francon, J., Nunez, J., and McManus, L. (1981).J. Cell Biol. 89, 680–683.
Luduena, R. F., Fellous, A., McManus, L. M., Jordan, M. A., and Nunez, J. (1984).J. Biol. Chem. 259, 12890–12898.
Mandelbaum-Shavit, F., Wolpert-DeFilippes, M. K., and Johns, D. G. (1976).Biochem. Biophys. Res. Commun. 72, 47–54.
Matus, A. (1994). InMicrotubules (Hyans, J. S., and Lloyd, C. W., eds.), Wiley-Liss, New York, pp. 155–166.
Okabe, S., and Hirokawa, N. (1989).Proc. Natl. Acad. Sci. USA 86, 4127–4131.
Prasad, V., Jordan, M. A., and Luduena, R. F. (1992).J. Protein Chem. 11, 509–515.
Roach, M. C., and Luduena, R. F. (1984).J. Biol. Chem. 259, 12063–12071.
Sandoval, I. V., and Vanderkerchove, J. S. (1981).J. Biol. Chem. 256, 8795–8800.
Sandoval, I. V., and Weber, K. (1980).J. Biol. Chem. 255, 8952–8954.
Sattilaro, R. F., Dentler, W. L., and LeCluyse, E. L. (1981).J. Cell Biol. 90, 467–473.
Vallee, R. B. (1980).Proc. Nat. Acad. Sci. USA 77, 3206–3210.
Witman, G. B., Cleveland, D. W., Weingarten, M. D., and Kirschner, M. W. (1976).Proc. Nat. Acad. Sci. USA 73, 4070–4074.
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Fellous, A., Prasad, V., Ohayon, R. et al. Removal of the projection domain of microtubule-associated protein 2 alters its interaction with tubulin. J Protein Chem 13, 381–391 (1994). https://doi.org/10.1007/BF01901694
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DOI: https://doi.org/10.1007/BF01901694