The spindle pole body component Spc97p interacts with the γ-tubulin of Saccharomyces cerevisiae and functions in microtubule organization and spindle pole body duplication

Michael Knop, Gislene Pereira, Silke Geissler, Katrin Grein, Elmar Schiebel

Research output: Contribution to journalArticle

143 Citations (Scopus)

Abstract

Previously, we have shown that the γ-tubulin Tub4p and the spindle pole body component Spc98p are involved in microtubule organization by the yeast microtubule organizing centre, the spindle pole body (SPB). In this paper we report the identification of SPC97 encoding an essential SPB component that is in association with the SPB substructures that organize the cytoplasmic and nuclear microtubules. Evidence is provided for a physical and functional interaction between Tub4p, Spc98p and Spc97p: first, temperature-sensitive spc97(ts) mutants are suppressed by high gene dosage of SPC98 or TUB4. Second, Spc97p interacts with Spc98p and Tub4p in the two-hybrid system. Finally, immunoprecipitation and fractionation studies revealed complexes containing Tub4p, Spc98p and Spc97p. Further support for a direct interaction of Tub4p, Spc98p and Spc97p comes from the toxicity of strong SPC97 overexpression which is suppressed by co-overexpression of TUB4 or SPC98. Analysis of temperature-sensitive spc97(ts) alleles revealed multiple spindle defects. While spc97-14 cells are either impaired in SPB separation or mitotic spindle formation, spc97-20 cells show an additional defect in SPB duplication. We discuss a model in which the Tub4p-Spc98p-Spc97p complex is part of the microtubule attachment site at the SPB.

Original languageEnglish (US)
Pages (from-to)1550-1564
Number of pages15
JournalEMBO Journal
Volume16
Issue number7
DOIs
StatePublished - Apr 1 1997

Fingerprint

Spindle Pole Bodies
Tubulin
Microtubules
Yeast
Saccharomyces cerevisiae
Poles
Microtubule-Organizing Center
Defects
Spindle Apparatus
Temperature
Gene Dosage
Fractionation
Hybrid systems
Immunoprecipitation
Toxicity
Genes
Yeasts
Alleles

Keywords

  • Microtubule organization
  • Spc97p
  • Spc98p
  • Spindle pole body
  • Tub4p
  • γ-tubulin

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

The spindle pole body component Spc97p interacts with the γ-tubulin of Saccharomyces cerevisiae and functions in microtubule organization and spindle pole body duplication. / Knop, Michael; Pereira, Gislene; Geissler, Silke; Grein, Katrin; Schiebel, Elmar.

In: EMBO Journal, Vol. 16, No. 7, 01.04.1997, p. 1550-1564.

Research output: Contribution to journalArticle

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