|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 109, Issue 1 165-177, Copyright © 1996 by Company of Biologists
JOURNAL ARTICLES |
H Masuda and T Shibata
Precursory Research for Embryonic Science and Technology (PRESTO), Research Development Corporation of Japan (JRDC).
The ability of the Schizosacchromyces pombe spindle pole body to nucleate microtubules is activated at the onset of mitosis for forming a mitotic spindle, but it is inactivated during interphase. We have previously developed an in vitro assay for studying the molecular mechanism of spindle pole body activation using permeabilized interphase S. pombe cells and Xenopus mitotic extracts. We have shown that the interphase spindle pole body is activated indirectly by p34cdc2 protein kinase in Xenopus mitotic extracts. In this study we examined the role of gamma-tubulin, a component of both interphase and mitotic spindle pole body, in formation of the microtubule nucleating complex at the mitotic spindle pole body. A polyclonal antibody specific to S. pombe gamma-tubulin inhibited both activation of the interphase spindle pole body and microtubule nucleation from the mitotic spindle pole body. Addition of bacterially expressed S. pombe gamma-tubulin or its amino-terminal fragments to Xenopus mitotic extracts inhibited spindle pole body activation. Affinity chromatography of partially fractionated Xenopus mitotic extracts with the amino-terminal fragment of S. pombe gamma-tubulin showed that fractions bound to the fragment supported the activation. The fractions did not contain Xenopus gamma-tubulin, showing that activation of the spindle pole body is not due to recruitment of Xenopus gamma-tubulin to the spindle pole body. The spindle pole body activation occurred in extracts depleted of p34cdc2 protein kinase or MAP kinase. The activity of the fractions bound to the fragment was inhibited by a protein kinase inhibitor, staurosporine. These results suggest that S. pombe gamma-tubulin is a component of the microtubule nucleating complex, and that the function of proteins that interact with gamma-tubulin is required for activation of the spindle pole body. We present possible models for the activation that convert the immature microtubule nucleating complex at interphase into the mature microtubule nucleating complex at mitosis.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
 |
|
  L. Cuschieri, R. Miller, and J. Vogel {gamma}-Tubulin Is Required for Proper Recruitment and Assembly of Kar9-Bim1 Complexes in Budding Yeast Mol. Biol. Cell, October 1, 2006; 17(10): 4420 - 4434. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Masuda, T. Toda, R. Miyamoto, T. Haraguchi, and Y. Hiraoka Modulation of Alp4 function in Schizosaccharomyces pombe induces novel phenotypes that imply distinct functions for nuclear and cytoplasmic {gamma}-tubulin complexes Genes Cells, April 1, 2006; 11(4): 319 - 336. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Ovechkina, P. Maddox, C. E. Oakley, X. Xiang, S. A. Osmani, E. D. Salmon, and B. R. Oakley Spindle Formation in Aspergillus Is Coupled to Tubulin Movement into the Nucleus Mol. Biol. Cell, May 1, 2003; 14(5): 2192 - 2200. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Takada, T. Shibata, Y. Hiraoka, and H. Masuda Identification of Ribonucleotide Reductase Protein R1 as an Activator of Microtubule Nucleation in Xenopus Egg Mitotic Extracts Mol. Biol. Cell, December 1, 2000; 11(12): 4173 - 4187. [Abstract] [Full Text]
|
|
|
|
 |
|
 Y.-c. Li, C.-r. Chen, and E. C. Chang Fission Yeast Ras1 Effector Scd1 Interacts With the Spindle and Affects Its Proper Formation Genetics, November 1, 2000; 156(3): 995 - 1004. [Abstract] [Full Text]
|
|
|
|
|
|
J. L. Paluh, E. Nogales, B. R. Oakley, K. McDonald, A. L. Pidoux, and W. Z. Cande A Mutation in gamma -Tubulin Alters Microtubule Dynamics and Organization and Is Synthetically Lethal with the Kinesin-like Protein Pkl1p Mol. Biol. Cell, April 1, 2000; 11(4): 1225 - 1239. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. Nakamura, H. Masuda, J. Horii, K.-i. Kuma, N. Yokoyama, T. Ohba, H. Nishitani, T. Miyata, M. Tanaka, and T. Nishimoto When Overexpressed, a Novel Centrosomal Protein, RanBPM, Causes Ectopic Microtubule Nucleation Similar to {gamma}-Tubulin J. Cell Biol., November 16, 1998; 143(4): 1041 - 1052. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Kimura, M. Hirano, R. Kobayashi, and T. Hirano Phosphorylation and Activation of 13S Condensin by Cdc2 in Vitro Science, October 16, 1998; 282(5388): 487 - 490. [Abstract] [Full Text]
|
|
|
|
|
|
A.-M. Tassin, C. Celati, M. Moudjou, and M. Bornens Characterization of the Human Homologue of the Yeast Spc98p and Its Association with {gamma}-Tubulin J. Cell Biol., May 4, 1998; 141(3): 689 - 701. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X He, M. Jones, M Winey, and S Sazer Mph1, a member of the Mps1-like family of dual specificity protein kinases, is required for the spindle checkpoint in S. pombe J. Cell Sci., January 6, 1998; 111(12): 1635 - 1647. [Abstract] [PDF]
|
|
|
|
|
|
M. Martin, S. Osmani, and B. Oakley The role of gamma-tubulin in mitotic spindle formation and cell cycle progression in Aspergillus nidulans J. Cell Sci., January 3, 1997; 110(5): 623 - 633. [Abstract] [PDF]
|
|
