Mutational analyses reveal a novel function of the nucleotide-binding domain of gamma-tubulin in the regulation of basal body biogenesis.
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.
Shang
Yuhua
Y
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Tsao
Che-Chia
CC
Gorovsky
Martin A
MA
D000818:Animals; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods; D005786:Gene Expression Regulation / Q000502:physiology*; D005998:Glycine / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D022101:Microtubule-Organizing Center / Q000502:physiology*; D008954:Models, Biological; D017354:Point Mutation; D017434:Protein Structure, Tertiary / Q000235:genetics* / Q000502:physiology; D016808:Tetrahymena thermophila; D014404:Tubulin / Q000235:genetics* / Q000378:metabolism
D014404:Tubulin; D005998:Glycine