Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells
Open Access
- 7 January 2002
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 156 (1), 87-100
- https://doi.org/10.1083/jcb.200108088
Abstract
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 α-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.Keywords
This publication has 48 references indexed in Scilit:
- Analysis of the Saccharomyces Spindle Pole by Matrix-assisted Laser Desorption/Ionization (MALDI) Mass SpectrometryThe Journal of cell biology, 1998
- Pericentrin and γ-Tubulin Form a Protein Complex and Are Organized into a Novel Lattice at the CentrosomeThe Journal of cell biology, 1998
- The Cell Center at 100Cell, 1997
- A Sequence Property Approach to Searching Protein DatabasesJournal of Molecular Biology, 1995
- Coiled-coil packing in spermine-induced tropomyosin crystals a comparative study of three formsJournal of Molecular Biology, 1994
- Heterogeneity of microtubule organizing center components as revealed by monoclonal antibodies to mammalian centrosomes and to nucleus‐associated bodies from DictyosteliumCell Motility, 1992
- Predicting Coiled Coils from Protein SequencesScience, 1991
- Expression of chicken lamin B2 in Escherichia coli: characterization of its structure, assembly, and molecular interactions.The Journal of cell biology, 1991
- 225‐Kilodalton phosphoprotein associated with mitotic centrosomes in sea urchin eggsCell Motility, 1989
- Is the centriole bound to the nuclear membrane?Nature, 1977