Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation

Nature. 1992 Mar 5;356(6364):80-3. doi: 10.1038/356080a0.

Abstract

gamma-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 microtubule organizing centres and a mutation in the gene encoding it produces a microtubuleless mitotic arrest in the filamentous fungus Aspergillus nidulans. Here we investigate the in vivo function of gamma-tubulin in mammalian cells using a synthetic peptide to generate a polyclonal antibody that binds to a highly conserved segment of gamma-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 gamma-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 gamma-tubulin is required for microtubule nucleation throughout the mammalian cell cycle.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Cell Cycle / drug effects
  • Cell Cycle / physiology*
  • Cells, Cultured
  • Chromosomes / metabolism
  • Cold Temperature
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Mice
  • Microtubules / metabolism*
  • Molecular Sequence Data
  • Nocodazole / pharmacology
  • Nuclear Proteins / metabolism*
  • Peptide Fragments / metabolism
  • Spindle Apparatus / metabolism*
  • Tubulin / metabolism*

Substances

  • Nuclear Proteins
  • Peptide Fragments
  • Tubulin
  • Nocodazole