The spindle pole body component Spc98p interacts with the gamma-tubulin-like Tub4p of Saccharomyces cerevisiae at the sites of microtubule attachment.
Tub4p is a novel tubulin found in Saccharomyces cerevisiae. It most resembles gamma-tubulin and, like it, is localized to the yeast microtubule organizing centre, the spindle pole body (SPB). In this paper we report the identification of SPC98 as a dosage-dependent suppressor of the conditional lethal tub4-1 allele. SPC98 encodes ... an SPB component of 98 kDa which is identical to the previously described 90 kDa SPB protein. Strong overexpression of SPC98 is toxic, causing cells to arrest with a large bud, defective microtubule structures, undivided nucleus and replicated DNA. The toxicity of SPC98 overexpression was relieved by co-overexpression of TUB4. Further evidence for an interaction between Tub4p and Spc98p came from the synthetic toxicity of tub4-1 and spc98-1 alleles, the dosage-dependent suppression of spc98-4 by TUB4, the binding of Tub4p to Spc98p in the two-hybrid system and the co-immunoprecipitation of Tub4p and Spc98p. In addition, Spc98-1p is defective in its interaction with Tub4p in the two-hybrid system. We suggest a model in which Tub4p and Spc98p form a complex involved in microtubule organization by the SPB.
Mesh Terms:
Amino Acid Sequence, Cell Cycle, Cell Survival, Cloning, Molecular, Fungal Proteins, Gene Expression Regulation, Fungal, Genes, Suppressor, Microscopy, Fluorescence, Mitotic Spindle Apparatus, Molecular Sequence Data, Phenotype, Plasmids, Promoter Regions, Genetic, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Temperature, Tubulin
Amino Acid Sequence, Cell Cycle, Cell Survival, Cloning, Molecular, Fungal Proteins, Gene Expression Regulation, Fungal, Genes, Suppressor, Microscopy, Fluorescence, Mitotic Spindle Apparatus, Molecular Sequence Data, Phenotype, Plasmids, Promoter Regions, Genetic, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Temperature, Tubulin
EMBO J.
Date: Aug. 01, 1996
PubMed ID: 8670895
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