The yeast spindle pole body component Spc72p interacts with Stu2p and is required for proper microtubule assembly.

The yeast spindle pole body component Spc72p interacts with Stu2p and is required for proper microtubule assembly.
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DOI:
10.1083/jcb.141.5.1169
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发表时间:
1998-06-01
影响因子:
7.8
通讯作者:
Huffaker, TC
Huffaker, TC
中科院分区:
生物学1区
文献类型:
--
作者:
Chen, XYP;Yin, HW;Huffaker, TC

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我们之前已经证明 Stu2p 是一种微管结合蛋白,也是酿酒酵母纺锤体极体 (SPB) 的组成部分。在这里,我们报告了 Spc72p 的鉴定,这是一种与 Stu2p 相互作用的蛋白质。 Stu2p 和 Spc72p 在双杂交系统中结合,可以从酵母提取物中进行免疫共沉淀。 Stu2p 和 Spc72p 也与自身相互作用,表明多聚体 Stu2p-Spc72p 复合物的可能性。 Spc72p 是 SPB 的重要组成部分,能够与预先存在的 SPB 结合,表明 Spc72p 的可溶性形式和 SPB 形式之间存在动态交换。与 Stu2p 不同,Spc72p 在体外不结合微管,并且在体内未观察到沿微管定位。温度敏感的 spc72 突变会导致 SPB 形态缺陷。此外,大多数spc72突变细胞缺乏细胞质微管;观察到的少数细胞质微管过长,其中一些未附着在 SPB 上。 spc72细胞能够复制和分离其SPB以形成双极纺锤体,但纺锤体伸长和染色体分离很少发生。染色体分离阻断不会阻止细胞周期;相反,spc72 细胞经历胞质分裂,产生包含多个 SPB 的非倍体细胞和多倍体细胞。
We have previously shown that Stu2p is a microtubule-binding protein and a component of the Saccharomyces cerevisiae spindle pole body (SPB). Here we report the identification of Spc72p, a protein that interacts with Stu2p. Stu2p and Spc72p associate in the two-hybrid system and can be coimmunoprecipitated from yeast extracts. Stu2p and Spc72p also interact with themselves, suggesting the possibility of a multimeric Stu2p-Spc72p complex. Spc72p is an essential component of the SPB and is able to associate with a preexisting SPB, indicating that there is a dynamic exchange between soluble and SPB forms of Spc72p. Unlike Stu2p, Spc72p does not bind microtubules in vitro, and was not observed to localize along microtubules in vivo. A temperature-sensitive spc72 mutation causes defects in SPB morphology. In addition, most spc72 mutant cells lack cytoplasmic microtubules; the few cytoplasmic microtubules that are observed are excessively long, and some of these are unattached to the SPB. spc72 cells are able to duplicate and separate their SPBs to form a bipolar spindle, but spindle elongation and chromosome segregation rarely occur. The chromosome segregation block does not arrest the cell cycle; instead, spc72 cells undergo cytokinesis, producing aploid cells and polyploid cells that contain multiple SPBs.
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影响因子: --
作者:
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