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PMID: 8707828 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Analysis of Tub4p, a yeast gamma-tubulin-like protein: implications for microtubule-organizing center function.

The Journal of cell biology ·Vol. 134 ·No. 2 ·1996-07-00 ·Pages 443-54

Marschall LG, Jeng RL, Mulholland J, Stearns T

Abstract

gamma-Tubulin is a conserved component of microtubule-organizing centers and is thought to be involved in microtubule nucleation. A recently discovered Saccharomyces cerevisiae gene (TUB4) encodes a tubulin that is related to, but divergent from, gamma-tubulins. TUB4 is essential for cell viability, and epitope-tagged Tub4 protein (Tub4p) is localized to the spindle pole body (Sobel, S.G., and M. Snyder. 1995.J. Cell Biol. 131:1775-1788). We have characterized the expression of TUB4, the association of Tub4p with the spindle pole body, and its role in microtubule organization. Tub4p is a minor protein in the cell, and expression of TUB4 is regulated in a cell cycle-dependent manner. Wild-type Tub4p is localized to the spindle pole body, and a Tub4p-green fluorescent protein fusion is able to associate with a preexisting spindle pole body, suggesting that there is dynamic exchange between cytoplasmic and spindle pole body forms of Tub4p. Perturbation of Tub4p function, either by conditional mutation or by depletion of the protein, results in spindle as well as spindle pole body defects, but does not eliminate the ability of microtubules to regrow from, or remain attached to, the spindle pole body. The spindle pole bodies in tub4 mutant cells duplicate but do not separate, resulting in a monopolar spindle. EM revealed that one spindle pole body of the duplicated pair appears to be defective for the nucleation of microtubules. These results offer insight into the role of gamma-tubulin in microtubule-organizing center function.

MeSH Terms
Animals Base Sequence DNA, Fungal Genes, Fungal Microtubules/physiology Molecular Sequence Data Mutation Phenotype Rabbits Saccharomyces cerevisiae/genetics,physiology Saccharomyces cerevisiae Proteins Spindle Apparatus/physiology Tubulin/genetics,physiology
Chemicals
DNA, Fungal Saccharomyces cerevisiae Proteins TUB4 protein, S cerevisiae Tubulin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Marschall L G
Department of Biological Sciences, Stanford University, California 94305-5020, USA.
Jeng R L
Mulholland J
Stearns T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.134.2.443
Published
1996-07-00
Pages
443-54
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120884
Subset
IM
Grants
NIGMS NIH HHS · GM 52022 · United States
External Links
PubMed source DOI: 10.1083/jcb.134.2.443 PMC Full Text View journal details
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