A gamma-tubulin that associates specifically with centrioles in HeLa cells and the basal body complex in Chlamydomonas.
gamma-Tubulin is a putative component of microtubule initiating material. To further explore its subcellular distribution in plant and animal cells, we have raised a polyclonal antibody, Rb27, directed towards a conserved region (EEFATEGTDRKDVFFY) of the gamma-tubulin molecule. Immunoblotting of cell protein extracts with Rb27 reveals a polypeptide band of M(r) 49 kD in HeLa and a 58 kD band in Chlamydomonas. Although these polypeptides are comparable in size to forms of gamma-tubulin detected previously in mammalian and plant protein extracts by other antibodies to gamma-tubulin, by immunofluorescence microscopy Rb27 gives localization patterns not previously seen. It localizes specifically with the centrioles in HeLa cells and with the basal body complex in Chlamydomonas. Other gamma-tubulin antibodies label pericentriolar material. Because of the similarities in the size of the polypeptides recognized by our and other gamma-tubulin antibodies, and a restricted co-localization with known microtubule-organizing centres in evolutionarily distant organisms, we propose that Rb27 recognizes a novel conserved gamma-tubulin isotype.
Dibbayawan
T P
TP
School of Biological Sciences, University of Sydney, Australia.
Harper
J D
JD
Elliott
J E
JE
Gunning
B E
BE
Marc
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002502:Centrioles / Q000737:chemistry*; D002696:Chlamydomonas / Q000737:chemistry* / Q000648:ultrastructure; D017124:Conserved Sequence; D006367:HeLa Cells; D006801:Humans; D007106:Immune Sera; D015151:Immunoblotting; D008969:Molecular Sequence Data; D014404:Tubulin / Q000032:analysis*
D007106:Immune Sera; D014404:Tubulin