Yang C H, Snyder M
Department of Biology, Yale University, New Haven, Connecticut 06511.
Mol Biol Cell. 1992 Nov;3(11):1259-67. doi: 10.1091/mbc.3.11.1259.
The formation and maintenance of the bipolar mitotic spindle apparatus require a complex and balanced interplay of several mechanisms, including the stabilization and separation of polar microtubules and the action of various microtubule motors. Nonmicrotubule elements are also present throughout the spindle apparatus and have been proposed to provide a structural support for the spindle. The Nuclear-Mitotic Apparatus protein (NuMA) is an abundant 240 kD protein that is present in the nucleus of interphase cells and concentrates in the polar regions of the spindle apparatus during mitosis. Sequence analysis indicates that NuMA possesses an unusually long alpha-helical central region characteristic of many filament forming proteins. In this report we demonstrate that microinjection of anti-NuMA antibodies into interphase and prophase cells results in a failure to form a mitotic spindle apparatus. Furthermore, injection of metaphase cells results in the collapse of the spindle apparatus into a monopolar microtubule array. These results identify for the first time a nontubulin component important for both the establishment and stabilization of the mitotic spindle apparatus in multicellular organisms. We suggest that nonmicrotubule structural components may be important for these processes.
双极有丝分裂纺锤体装置的形成和维持需要多种机制的复杂且平衡的相互作用,包括极微管的稳定和分离以及各种微管马达的作用。非微管成分也存在于整个纺锤体装置中,并被认为可为纺锤体提供结构支撑。核有丝分裂装置蛋白(NuMA)是一种丰富的240 kD蛋白,存在于间期细胞的细胞核中,并在有丝分裂期间集中于纺锤体装置的极区。序列分析表明,NuMA具有许多丝状形成蛋白特有的异常长的α螺旋中央区域。在本报告中,我们证明向间期和前期细胞显微注射抗NuMA抗体导致无法形成有丝分裂纺锤体装置。此外,向中期细胞注射会导致纺锤体装置塌陷为单极微管阵列。这些结果首次确定了一种对多细胞生物中有丝分裂纺锤体装置的建立和稳定都很重要的非微管蛋白成分。我们认为非微管结构成分可能对这些过程很重要。