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Published 7 January 2002. doi:10.1083/jcb.200108088
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© The Rockefeller University Press, 0021-9525/2002/1/87 $5.00
The Journal of Cell Biology, Volume 156, Number 1, January 7, 2002 87-100

Article

 Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells

Toshiro Ohta1, Russell Essner1, Jung-Hwa Ryu1, Robert E. Palazzo2, Yumi Uetake1 and Ryoko Kuriyama1

1 Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455
2 Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045

Address correspondence to Ryoko Kuriyama, Department of Genetics, Cell Biology, and Development, 6-160 Jackson Hall, 321 Church St. S.E., University of Minnesota, Minneapolis, MN 55455. Tel.: (612) 624-0471. Fax: (612) 626-6140. E-mail: ryoko{at}lenti.med.umn.edu

By using monoclonal antibodies raised against isolated clam centrosomes, we have identified a novel 135-kD centrosomal protein (Cep135), present in a wide range of organisms. Cep135 is located at the centrosome throughout the cell cycle, and localization is independent of the microtubule network. It distributes throughout the centrosomal area in association with the electron-dense material surrounding centrioles. Sequence analysis of cDNA isolated from CHO cells predicted a protein of 1,145–amino acid residues with extensive {alpha}-helical domains. Expression of a series of deletion constructs revealed the presence of three independent centrosome-targeting domains. Overexpression of Cep135 resulted in the accumulation of unique whorl-like particles in both the centrosome and the cytoplasm. Although their size, shape, and number varied according to the level of protein expression, these whorls were composed of parallel dense lines arranged in a 6-nm space. Altered levels of Cep135 by protein overexpression and/or suppression of endogenous Cep135 by RNA interference caused disorganization of interphase and mitotic spindle microtubules. Thus, Cep135 may play an important role in the centrosomal function of organizing microtubules in mammalian cells.

Key Words: centrosome; pericentriolar material; mitotic spindle; coiled-coil protein; RNAi


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