Department of Biochemistry and Biophysics, UCSF, San Francisco, California 94158, USA.
Cytoskeleton (Hoboken). 2010 Aug;67(8):504-18. doi: 10.1002/cm.20462.
The functional role of centrioles or basal bodies in mitotic spindle assembly and function is currently unclear. Although supernumerary centrioles have been associated with multipolar spindles in cancer cells, suggesting centriole number might dictate spindle polarity, bipolar spindles are able to assemble in the complete absence of centrioles, suggesting a level of centriole-independence in the spindle assembly pathway. In this report we perturb centriole number using mutations in Chlamydomonas reinhardtii, and measure the response of the mitotic spindle to these perturbations in centriole number. Although altered centriole number increased the frequency of monopolar and multipolar spindles, the majority of spindles remained bipolar regardless of the centriole number. But even when spindles were bipolar, abnormal centriole numbers led to asymmetries in tubulin distribution, half-spindle length and spindle pole focus. Half spindle length correlated directly with number of centrioles at a pole, such that an imbalance in centriole number between the two poles of a bipolar spindle correlated with increased asymmetry between half spindle lengths. These results are consistent with centrioles playing an active role in regulating mitotic spindle length. Mutants with centriole number alteration also show increased cytokinesis defects, but these do not correlate with centriole number in the dividing cell and may therefore reflect downstream consequences of defects in preceding cell divisions.
中心体或基体在有丝分裂纺锤体组装和功能中的作用目前尚不清楚。虽然多余的中心体与癌细胞中的多极纺锤体有关,表明中心体数量可能决定纺锤体极性,但在没有中心体的情况下也能组装出两极纺锤体,这表明在纺锤体组装途径中存在一定程度的中心体独立性。在本报告中,我们使用莱茵衣藻中的突变来改变中心体的数量,并测量有丝分裂纺锤体对这些中心体数量变化的反应。尽管改变中心体的数量增加了单极和多极纺锤体的频率,但大多数纺锤体仍然是两极的,而不管中心体的数量如何。但是,即使纺锤体是两极的,异常的中心体数量也会导致微管蛋白分布、半纺锤体长度和纺锤体极焦点的不对称。半纺锤体长度与极上的中心体数量直接相关,因此,两极上的中心体数量不平衡与半纺锤体长度之间的增加的不对称性相关。这些结果与中心体在调节有丝分裂纺锤体长度方面发挥积极作用是一致的。中心体数量改变的突变体也表现出更多的胞质分裂缺陷,但这些缺陷与分裂细胞中的中心体数量无关,因此可能反映了前几次细胞分裂缺陷的下游后果。