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1997), and fungi (Rout and Kilmartin, 1990), have been used. All these procedures have provided a necessary step for the biochemical, structural, and functional characterization of the centrosome. The aim of this chapter is to provide a brief overview of these methods and to discuss the results and limitations of these approaches. It will then deal with the more recent use of evolutionary conservation to identify essential centrosomal components. Finally, we discuss what the past can teach us for future studies of this unique organelle.
Academic Press, New York. , Marty, M. , and Karsenti, E. (1987). Structural and chemical characterization of isolated centrosomes. Cell Motil. Cytoskel. 8,238-249. , and Celati, C. (1989). The cortical microfilament system of lymphoblasts displays a periodic oscillatory activity in the absence of microtubules: Implication for cell polarity. J. Cell Biol. 109, 1071-1083. , Ferguson, J. , and Crocker, P. R. (1996a). Molecular characterization of Ninein, a new coiled-coil protein of the centrosome that binds spindle microtubules at mitosis.
Functional Assays of Isolated Centrosomes A. Microtubule Nucleating Activity B. Centrosome Biogenesis C. Comments Prospects A. What Does the Past Teach Us? B. Studying Centrosome Functions in Vivo References ME'IHUUS IN CELL BIOLOGY. VOL. h l Copynght 6 1999 by Acadermc Prrra. 110 ~ 13 Michel Bornens and Mohammed Moudjou 14 I. Introduction The centrosome is the microtubule organizing center of animal cells. During cell division, the duplication and segregation of centrosomes dictate the bipolarity of the mitotic spindle and thus determine the plane of the cleavage furrow.