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γ-Tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation

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Abstract

γ-TUBULIN is a newly identified member of the tubulin family whose sequence is highly conserved from yeast to man. This minor microtubule protein is localized to the microtuble organizing centres1–3 and a mutation in the gene encoding it produces a microtubuleless mitotic arrest in the filamentous fungus Aspergillus nidulans4,5. Here we investigate the in vivo function of γ-tubulin in mammalian cells using a synthetic peptide to generate a polyclonal antibody that binds to a highly conserved segment of γ-tubulin. After microinjection into cultured mammalian cells, immunofluorescence localization revealed that this antibody binds to native centrosomes at all phases of the cell cycle. In the presence of the γ-tubulin antibody, microtubules fail to regrow into cytoplasmic arrays after depolymerization induced by nocodazole or cold. Furthermore, cells injected immediately before or during mitosis fail to assemble a functional spindle. Thus in vivo γ-tubulin is required for microtubule nucleation throughout the mammalian cell cycle.

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Author notes

  1. Don W. Cleveland: Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Emory University School of Medicine, Atlanta, Georgia, 30322, USA

    Harish C. Joshi, Monica J. Palacios, Leemore McNamara & Don W. Cleveland

Authors
  1. Harish C. Joshi
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  2. Monica J. Palacios
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  3. Leemore McNamara
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  4. Don W. Cleveland
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Joshi, H., Palacios, M., McNamara, L. et al. γ-Tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. Nature 356, 80–83 (1992). https://doi.org/10.1038/356080a0

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  • DOI: https://doi.org/10.1038/356080a0

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