In mammals, γ-tubulin, a key protein in microtubule nucleation, is encoded by two genes, TUBG1 and TUBG2. Human TUBG1 and TUBG2 mRNA are expressed ubiquitously and predominantly in preimplantation embryos and the brain, respectively, but specific detection of γ-tubulin2 protein expression is difficult due to their high sequence similarity. Here, we describe a protocol for differential detection of two human γ-tubulins by western blotting. In several cancer cell lines and the brain, expression of γ-tubulin2 along with γ-tubulin1 and a novel TUBG2 splice variant are identified. Contribution of ectopically expressed γ-tubulin2 in cancer growth was determined by depletion of γ-tubulin2.
Keywords: cancer; centrosome; γ-tubulin.
© 2016 Federation of European Biochemical Societies.