Cooke C A, Heck M M, Earnshaw W C
Department of Cell Biology and Anatomy, Johns Hopkins School of Medicine, Baltimore, Maryland 21205.
J Cell Biol. 1987 Nov;105(5):2053-67. doi: 10.1083/jcb.105.5.2053.
We describe a novel set of polypeptide antigens that shows a dramatic change in structural localization during mitosis. Through metaphase these antigens define a new chromosomal substructure that is located between the sister chromatids. Because the antigens are concentrated in the pericentromeric region, we have provisionally termed them the INCENPs (inner centromere proteins). The INCENPs (two polypeptides of 155 and 135 kD) were identified with a monoclonal antibody that was raised against the bulk proteins of the mitotic chromosome scaffold fraction. These two polypeptides are the most tightly bound chromosomal proteins known. When scaffolds are prepared, 100% of the detectable INCENPs remain scaffold associated. We were therefore unprepared for the fate of the INCENPs at anaphase. As the sister chromatids separate, the INCENPs dissociate fully from them, remaining behind at the metaphase plate as the chromatids migrate to the spindle poles. During anaphase the INCENPs are found on coarse fibers in the central spindle, and also in close apposition to the cell membrane in the region of the forming contractile ring. During telophase, the INCENPs gradually become focused onto the forming midbody, together with which they are ultimately discarded. Several possible in vivo roles for the INCENPs are suggested by these data: regulation of sister chromatid pairing, stabilization of the plane of cleavage, and separation of spindle poles at anaphase.
我们描述了一组新型的多肽抗原,它们在有丝分裂期间的结构定位发生了显著变化。在中期,这些抗原定义了一种位于姐妹染色单体之间的新的染色体亚结构。由于这些抗原集中在着丝粒周围区域,我们暂时将它们称为着丝粒内蛋白(INCENPs)。INCENPs(155kD和135kD的两种多肽)是用一种针对有丝分裂染色体支架部分的整体蛋白产生的单克隆抗体鉴定出来的。这两种多肽是已知的与染色体结合最紧密的蛋白质。制备支架时,100%可检测到的INCENPs仍与支架相关。因此,我们对后期INCENPs的命运毫无准备。随着姐妹染色单体分离,INCENPs与它们完全解离,当染色单体向纺锤体极迁移时,INCENPs留在中期板处。在后期,INCENPs出现在中央纺锤体的粗纤维上,也紧密附着在形成收缩环区域的细胞膜上。在末期,INCENPs逐渐集中到正在形成的中间体上,并最终与之一起被丢弃。这些数据提示了INCENPs在体内的几种可能作用:姐妹染色单体配对的调节、分裂平面的稳定以及后期纺锤体极的分离。