Tousson A, Zeng C, Brinkley B R, Valdivia M M
Department of Cell Biology, University of Alabama, Birmingham, 35294.
J Cell Biol. 1991 Feb;112(3):427-40. doi: 10.1083/jcb.112.3.427.
A novel protein has been identified which may serve a key function in nucleating spindle microtubule growth in mitosis. This protein, called centrophilin, is sequentially relocated from the centromeres to the centrosomes to the midbody in a manner dependent on the mitotic phase. Centrophilin was initially detected by immunofluorescence with a monoclonal, primate-specific antibody (2D3) raised against kinetochore-enriched chromosome extract from HeLa cells (Valdivia, M. M., and B. R. Brinkley. 1985. J. Cell Biol. 101:1124-1134). Centrophilin forms prominent crescents at the poles of the metaphase spindle, gradually diminishes during anaphase, and bands the equatorial ends of midbody microtubules in telophase. The formation and breakdown of the spindle and midbody correlates in time and space with the aggregation and disaggregation of centrophilin foci. Immunogold EM reveals that centrophilin is a major component of pericentriolar material in metaphase. During recovery from microtubule inhibition, centrophilin foci act as nucleation sites for the assembly of spindle tubules. The 2D3 probe recognizes two high molecular mass polypeptides, 180 and 210 kD, on immunoblots of whole HeLa cell extract. Taken together, these data and the available literature on microtubule dynamics point inevitably to a singular model for control of spindle tubule turnover.
已鉴定出一种新蛋白质,它可能在有丝分裂中纺锤体微管生长的成核过程中发挥关键作用。这种名为着丝粒亲和蛋白的蛋白质,会根据有丝分裂阶段,依次从着丝粒重新定位到中心体,再到中间体。着丝粒亲和蛋白最初是通过免疫荧光法检测到的,使用的是一种针对从HeLa细胞富含动粒的染色体提取物产生的单克隆、灵长类特异性抗体(2D3)(瓦尔迪维亚,M.M.,和B.R.布林克利。1985年。《细胞生物学杂志》101:1124 - 1134)。着丝粒亲和蛋白在中期纺锤体的两极形成明显的新月形,在后期逐渐减少,并在末期环绕中间体微管的赤道端。纺锤体和中间体的形成与分解在时间和空间上与着丝粒亲和蛋白焦点的聚集和解聚相关。免疫金电子显微镜显示,着丝粒亲和蛋白是中期中心粒周围物质的主要成分。在从微管抑制中恢复期间,着丝粒亲和蛋白焦点充当纺锤体微管组装的成核位点。2D3探针在整个HeLa细胞提取物的免疫印迹上识别出两种高分子量多肽,180 kD和210 kD。综上所述,这些数据以及关于微管动力学的现有文献不可避免地指向一个控制纺锤体微管周转的单一模型。