Characterization of γ-tubulin complexes in Aspergillus nidulans and detection of putative γ-tubulin interacting proteins

Tomohiro Akashi, Yisang Yoon, Berl R. Oakley

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

γ-Tubulin is central to the nucleation of microtubule assembly in vivo. Although it is most obviously located at microtubule organizing centers, it is also found in soluble cytoplasmic complexes. Characterizing these complexes and identifying proteins that interact with γ-tubulin in vivo will be necessary if γ-tubulin function is to be understood fully. We have begun to investigate soluble complexes of γ-tubulin in Aspergillus nidulans, the organism in which γ-tubulin was discovered and in which a great deal of genetic and molecular genetic analysis of γ-tubulin has been carried out. We find that approximately 32% of the γ-tubulin in A. nidulans is soluble. Sucrose density gradients revealed that the soluble γ-tubulin is in 8-20S complexes with little or no monomeric γ-tubulin present. In the presence of 0.5 M KCl, the average size of the complexes decreased and a peak was present between 4S and 11S. Cross-linking experiments with a zero-length cross- linker suggest that γ-tubulin in isolated nuclei and in intact hyphae interacts physically with three proteins with molecular weights of approximately 105, 95, and 80 kDa.

Original languageEnglish (US)
Pages (from-to)149-158
Number of pages10
JournalCell Motility and the Cytoskeleton
Volume37
Issue number2
DOIs
StatePublished - Jun 26 1997
Externally publishedYes

Keywords

  • Microtubule organizing centers
  • Microtubules
  • Spindle pole bodies

ASJC Scopus subject areas

  • Structural Biology
  • Cell Biology

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