Hannak E, Kirkham M, Hyman A A, Oegema K
Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
J Cell Biol. 2001 Dec 24;155(7):1109-16. doi: 10.1083/jcb.200108051. Epub 2001 Dec 17.
Centrosomes mature as cells enter mitosis, accumulating gamma-tubulin and other pericentriolar material (PCM) components. This occurs concomitant with an increase in the number of centrosomally organized microtubules (MTs). Here, we use RNA-mediated interference (RNAi) to examine the role of the aurora-A kinase, AIR-1, during centrosome maturation in Caenorhabditis elegans. In air-1(RNAi) embryos, centrosomes separate normally, an event that occurs before maturation in C. elegans. After nuclear envelope breakdown, the separated centrosomes collapse together, and spindle assembly fails. In mitotic air-1(RNAi) embryos, centrosomal alpha-tubulin fluorescence intensity accumulates to only 40% of wild-type levels, suggesting a defect in the maturation process. Consistent with this hypothesis, we find that AIR-1 is required for the increase in centrosomal gamma-tubulin and two other PCM components, ZYG-9 and CeGrip, as embryos enter mitosis. Furthermore, the AIR-1-dependent increase in centrosomal gamma-tubulin does not require MTs. These results suggest that aurora-A kinases are required to execute a MT-independent pathway for the recruitment of PCM during centrosome maturation.
随着细胞进入有丝分裂,中心体逐渐成熟,积累γ-微管蛋白和其他中心粒外周物质(PCM)成分。这一过程伴随着中心体组织的微管(MTs)数量的增加。在这里,我们利用RNA介导的干扰(RNAi)技术来研究秀丽隐杆线虫中心体成熟过程中极光激酶A(aurora-A kinase),即AIR-1的作用。在进行air-1(RNAi)处理的胚胎中,中心体正常分离,这一事件发生在秀丽隐杆线虫中心体成熟之前。核膜破裂后,分离的中心体坍塌在一起,纺锤体组装失败。在有丝分裂的air-1(RNAi)胚胎中,中心体α-微管蛋白荧光强度仅积累到野生型水平的40%,这表明在成熟过程中存在缺陷。与这一假设一致,我们发现当胚胎进入有丝分裂时,AIR-1对于中心体γ-微管蛋白以及另外两种PCM成分ZYG-9和CeGrip的增加是必需的。此外,中心体γ-微管蛋白依赖于AIR-1的增加并不需要微管。这些结果表明,在中心体成熟过程中,极光激酶A对于执行一条不依赖微管的PCM招募途径是必需的。