Mutational analyses reveal a novel function of the nucleotide-binding domain of gamma-tubulin in the regulation of basal body biogenesis

J Cell Biol. 2005 Dec 19;171(6):1035-44. doi: 10.1083/jcb.200508184. Epub 2005 Dec 12.

Abstract

We have used in vitro mutagenesis and gene replacement to study the function of the nucleotide-binding domain (NBD) of gamma-tubulin in Tetrahymena thermophila. In this study, we show that the NBD has an essential function and that point mutations in two conserved residues lead to over-production and mislocalization of basal body (BB) assembly. These results, coupled with previous studies (Dammermann, A., T. Muller-Reichert, L. Pelletier, B. Habermann, A. Desai, and K. Oegema. 2004. Dev. Cell. 7:815-829; La Terra, S., C.N. English, P. Hergert, B.F. McEwen, G. Sluder, and A. Khodjakov. 2005. J. Cell Biol. 168:713-722), suggest that to achieve the precise temporal and spatial regulation of BB/centriole assembly, the initiation activity of gamma-tubulin is normally suppressed by a negative regulatory mechanism that acts through its NBD.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Cycle / genetics
  • Cell Cycle / physiology
  • Cytoplasm / metabolism
  • DNA Mutational Analysis / methods
  • Gene Expression Regulation / physiology*
  • Glycine / genetics
  • Glycine / metabolism
  • Immunohistochemistry
  • Microtubule-Organizing Center / physiology*
  • Models, Biological
  • Point Mutation
  • Protein Structure, Tertiary / genetics*
  • Protein Structure, Tertiary / physiology
  • Tetrahymena thermophila
  • Tubulin / genetics*
  • Tubulin / metabolism

Substances

  • Tubulin
  • Glycine