Vérollet Christel, Colombié Nathalie, Daubon Thomas, Bourbon Henri-Marc, Wright Michel, Raynaud-Messina Brigitte
Centre de Recherche en Pharmacologie, Santé, UMR 2587, Centre National de la Recherche Scientifique-Pierre Fabre, Institut de Sciences et Technologies du Médicament de Toulouse, 31432 Toulouse, Cedex 4, France.
J Cell Biol. 2006 Feb 13;172(4):517-28. doi: 10.1083/jcb.200511071.
In metazoans, gamma-tubulin acts within two main complexes, gamma-tubulin small complexes (gamma-TuSCs) and gamma-tubulin ring complexes (gamma-TuRCs). In higher eukaryotes, it is assumed that microtubule nucleation at the centrosome depends on gamma-TuRCs, but the role of gamma-TuRC components remains undefined. For the first time, we analyzed the function of all four gamma-TuRC-specific subunits in Drosophila melanogaster: Dgrip75, Dgrip128, Dgrip163, and Dgp71WD. Grip-motif proteins, but not Dgp71WD, appear to be required for gamma-TuRC assembly. Individual depletion of gamma-TuRC components, in cultured cells and in vivo, induces mitotic delay and abnormal spindles. Surprisingly, gamma-TuSCs are recruited to the centrosomes. These defects are less severe than those resulting from the inhibition of gamma-TuSC components and do not appear critical for viability. Simultaneous cosilencing of all gamma-TuRC proteins leads to stronger phenotypes and partial recruitment of gamma-TuSC. In conclusion, gamma-TuRCs are required for assembly of fully functional spindles, but we suggest that gamma-TuSC could be targeted to the centrosomes, which is where basic microtubule assembly activities are maintained.
在多细胞动物中,γ-微管蛋白在两个主要复合物中起作用,即γ-微管蛋白小复合物(γ-TuSCs)和γ-微管蛋白环复合物(γ-TuRCs)。在高等真核生物中,人们认为中心体处的微管成核依赖于γ-TuRCs,但γ-TuRC 组分的作用仍不明确。我们首次分析了果蝇中所有四种γ-TuRC特异性亚基的功能:Dgrip75、Dgrip128、Dgrip163和Dgp71WD。γ-TuRC组装似乎需要具有Grip基序的蛋白,但不需要Dgp71WD。在培养细胞和体内对γ-TuRC组分进行单独敲减会诱导有丝分裂延迟和纺锤体异常。令人惊讶的是,γ-TuSCs会被招募到中心体。这些缺陷不如抑制γ-TuSC组分所导致的缺陷严重,且对生存力似乎并非至关重要。同时共沉默所有γ-TuRC蛋白会导致更强的表型和γ-TuSC的部分招募。总之,功能性纺锤体的组装需要γ-TuRCs,但我们认为γ-TuSCs可能会被靶向到中心体,而中心体是维持基本微管组装活性的地方。