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酿酒酵母的CDC20基因产物是一种β-转导蛋白同源物,是微管依赖性细胞过程的一个子集所必需的。

The CDC20 gene product of Saccharomyces cerevisiae, a beta-transducin homolog, is required for a subset of microtubule-dependent cellular processes.

作者信息

Sethi N, Monteagudo M C, Koshland D, Hogan E, Burke D J

机构信息

Department of Biology, University of Virginia, Charlottesville 22901.

出版信息

Mol Cell Biol. 1991 Nov;11(11):5592-602. doi: 10.1128/mcb.11.11.5592-5602.1991.

Abstract

Previous analysis of cdc20 mutants of the yeast Saccharomyces cerevisiae suggests that the CDC20 gene product (Cdc20p) is required for two microtubule-dependent processes, nuclear movements prior to anaphase and chromosome separation. Here we report that cdc20 mutants are defective for a third microtubule-mediated event, nuclear fusion during mating of G1 cells, but appear normal for a fourth microtubule-dependent process, nuclear migration after DNA replication. Therefore, Cdc20p is required for a subset of microtubule-dependent processes and functions at multiple stages in the life cycle. Consistent with this interpretation, we find that cdc20 cells arrested by alpha-factor or at the restrictive temperature accumulate anomalous microtubule structures, as detected by indirect immunofluorescence. The anomalous microtubule staining patterns are due to cdc20 because intragenic revertants that revert the temperature sensitivity have normal microtubule morphologies. cdc20 mutants have a sevenfold increase in the intensity of antitubulin fluorescence in intranuclear spindles compared with spindles from wild-type cells, yet the total amount of tubulin is indistinguishable by Western immunoblot analysis. This result suggests that Cdc20p modulates microtubule structure in wild-type cells either by promoting microtubule disassembly or by altering the surface of the microtubules. Finally, we cloned and sequenced CDC20 and show that it encodes a member of a family of proteins that share homology to the beta subunit of transducin.

摘要

先前对酿酒酵母cdc20突变体的分析表明,CDC20基因产物(Cdc20p)参与两个依赖微管的过程,即后期之前的核运动和染色体分离。在此我们报道,cdc20突变体在第三个由微管介导的事件(G1期细胞交配时的核融合)中存在缺陷,但在第四个依赖微管的过程(DNA复制后的核迁移)中表现正常。因此,Cdc20p参与了部分依赖微管的过程,并在生命周期的多个阶段发挥作用。与这一解释相符的是,我们发现,通过间接免疫荧光检测,被α因子阻断或在限制温度下的cdc20细胞积累了异常的微管结构。异常的微管染色模式是由cdc20导致的,因为恢复温度敏感性的基因内回复突变体具有正常的微管形态。与野生型细胞纺锤体相比,cdc20突变体的核内纺锤体中抗微管蛋白荧光强度增加了7倍,但通过蛋白质免疫印迹分析,微管蛋白的总量并无差异。这一结果表明,Cdc20p在野生型细胞中通过促进微管解聚或改变微管表面来调节微管结构。最后,我们克隆并测序了CDC20,结果显示它编码一种与转导素β亚基具有同源性的蛋白质家族成员。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c46/361930/bdbcc36ad8f2/molcellb00035-0220-a.jpg

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