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Studies on molecular mechanisms of division furrow formation in Tetrahymena

机译:四膜虫分沟形成的分子机理研究

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摘要

Employment of mutants which have the defects in cell division makes it possible to elucidate the molecular mechanisms of cell division. I used cdaAl of a temperature-sensitive Tetrahymena mutant having a defect in the formation of fission zone where contractile ring is formed. I expected that the product of cdaA locus is a factor required for the formation of fission zone. In this study, I identified a protein showing a difference in electrophoretic mobility between wild-type and cdaAl cell as a possible gene product of cdaA locus. The product had a product of cdaA locus. The product had a molecular weight mass of 85,000, isoelectric point of 4.7, so that it was named p85. Expression of either wild-type p85 or cdaA p85 coincided with expression of respective phenotype in the descendants derived from the cross between cdaAl and wild-type cells, which strongly suggested that p85 is indeed a cdaA gene product. Furthermore, immunofluorescence staining demonstrated that specific localization of p85 occurred at equatorial basal bodies at the time just before the formation of fission zone which should be formed in front of the equatorial basal bodies. Such a localization must be prerequisite for cell division because the localization failed to occur in cdaAl cell at the restrictive temperature. It was found that contractile ring microfilaments ran along the basal bodies where p85 was localized. The fact suggested that p85 plays a important role in formations of both fission zone and contractile ring. Time and place of polymerization of contractile ring microfilaments were investigated in known-aged Tetrahymena cells by microinjecting skeletal muscle actin which might act as a functionally-incompatible analogue of Tetrahymena actin. The results suggested that the polymerization of contractile ring microfilaments occurred at cell equator in a 17-minutes-period before the onset of cell division. The polymerization of contractiloe ring microfilaments coincided with the fisson zone formation, and both events were only slightly preceded by the equatorial localization of p85. These facts lead a notion that p85 localization at equatorial basal bodies triggers both formation of fission zone and polymerization of contractile ring microfilaments, and that p85 provides the polymerizatiion nuclei for Tetrahymena action. It can be said that p85 identified as the gene product of cdaA locus is a crucial determinant factor for the time and place of division furrow constriction.
机译:使用在细胞分裂中具有缺陷的突变体使得有可能阐明细胞分裂的分子机制。我使用了对温度敏感的四膜虫突变体的cdaA1,该变体在形成收缩环的裂变区的形成中存在缺陷。我希望cdaA位点的产物是形成裂变区所需的一个因素。在这项研究中,我鉴定了一种蛋白质,该蛋白质在野生型和cdaA1细胞之间显示出电泳迁移率差异,是cdaA基因座的可能基因产物。该产物具有cdaA基因座的产物。该产物的分子量为85,000,等电点为4.7,因此命名为p85。野生型p85或cdaA p85的表达与来自cdaA1和野生型细胞之间杂交的后代中各自表型的表达相吻合,这强烈表明p85确实是cdaA基因产物。此外,免疫荧光染色表明p85的特定定位发生在裂变区形成之前的时间,该裂变区应该在赤道基体的前面形成。这种定位必须是细胞分裂的前提条件,因为在限制温度下cdaAl电池无法进行定位。发现收缩环微丝沿着p85定位的基体。事实表明,p85在裂变区和收缩环的形成中都起着重要作用。通过显微注射骨骼肌肌动蛋白来研究在已知年龄的四膜膜细胞中收缩环微丝聚合的时间和地点,它可能是四膜膜肌动蛋白的功能不相容的类似物。结果表明,收缩环微丝的聚合发生在细胞分裂开始之前的17分钟内的细胞赤道。收缩环微丝的聚合与费森区的形成相吻合,并且这两个事件仅在p85的赤道定位之前发生。这些事实导致一个观念,即p85定位在赤道基体上会触发裂变区的形成和收缩环微丝的聚合,并且p85为四膜虫的作用提供了聚合核。可以说,被鉴定为cdaA基因座基因产物的p85是决定犁沟收缩的时间和地点的关键决定因素。

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