Lüders Jens, Patel Urvashi K, Stearns Tim
Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA.
Nat Cell Biol. 2006 Feb;8(2):137-47. doi: 10.1038/ncb1349. Epub 2005 Dec 25.
The gamma-tubulin ring complex (gammaTuRC) is a large multi-protein complex that is required for microtubule nucleation from the centrosome. Here, we show that the GCP-WD protein (originally named NEDD1) is the orthologue of the Drosophila Dgrip71WD protein, and is a subunit of the human gammaTuRC. GCP-WD has the properties of an attachment factor for the gammaTuRC: depletion or inhibition of GCP-WD results in loss of the gammaTuRC from the centrosome, abolishing centrosomal microtubule nucleation, although the gammaTuRC is intact and able to bind to microtubules. GCP-WD depletion also blocks mitotic chromatin-mediated microtubule nucleation, resulting in failure of spindle assembly. Mitotic phosphorylation of GCP-WD is required for association of gamma-tubulin with the spindle, separately from association with the centrosome. Our results indicate that GCP-WD broadly mediates targeting of the gammaTuRC to sites of microtubule nucleation and to the mitotic spindle, which is essential for spindle formation.
γ-微管蛋白环复合物(γTuRC)是一种大型多蛋白复合物,是中心体微管成核所必需的。在此,我们表明GCP-WD蛋白(最初命名为NEDD1)是果蝇Dgrip71WD蛋白的直系同源物,并且是人类γTuRC的一个亚基。GCP-WD具有γTuRC附着因子的特性:GCP-WD的缺失或抑制会导致γTuRC从中心体丢失,消除中心体微管成核,尽管γTuRC是完整的并且能够与微管结合。GCP-WD的缺失还会阻断有丝分裂染色质介导的微管成核,导致纺锤体组装失败。GCP-WD的有丝分裂磷酸化是γ-微管蛋白与纺锤体结合所必需的,这与它和中心体的结合是分开的。我们的结果表明,GCP-WD广泛介导γTuRC靶向微管成核位点和有丝分裂纺锤体,这对纺锤体形成至关重要。