Foreman P K, Davis R W
Department of Biochemistry, Stanford University School of Medicine, California 94305, USA.
Genetics. 1996 Dec;144(4):1387-97. doi: 10.1093/genetics/144.4.1387.
To identify new gene products involved in chromosome segregation, we isolated Saccharomyces cerevisiae mutants that require centromere binding factor I (Cbf1p) for viability. One Cbf1p-dependent mutant (denoted cdp1-1) was selected for further analysis. The CDP1 gene encodes a novel 125-kD protein that is notably similar to previously identified mouse, human and Caenorhabditis elegans proteins. CDP1 delta and cdp1-1 mutant cells were temperature sensitive for growth. At the permissive temperature, cdp1-1 and cdp1 delta cells lost chromosomes at a frequencies approximately 20-fold and approximately 110-fold higher than wild-type cells, respectively. These mutants also displayed unusually long and numerous bundles of cytoplasmic microtubules as revealed by immunofluorescent staining. In addition, we occasionally observed improperly oriented mitotic spindles, residing entirely within one of the cells. Presumably as a result of undergoing nuclear division with improperly oriented spindles, a large percentage of cdp1 cells had accumulated multiple nuclei. While cdp1 mutant cells were hypersensitive to the microtubule-disrupting compound thiabendazole, they showed increased resistance to the closely related compound benomyl relative to wild-type cells. Taken together, these results suggest that Cdp1p plays a role in governing tubulin dynamics within the cell and may interact directly with microtubules or tubulin.
为了鉴定参与染色体分离的新基因产物,我们分离了酿酒酵母突变体,这些突变体需要着丝粒结合因子I(Cbf1p)才能存活。选择了一个依赖Cbf1p的突变体(命名为cdp1-1)进行进一步分析。CDP1基因编码一种新的125-kD蛋白,该蛋白与先前鉴定的小鼠、人类和秀丽隐杆线虫的蛋白显著相似。CDP1δ和cdp1-1突变体细胞的生长对温度敏感。在允许温度下,cdp1-1和cdp1δ细胞丢失染色体的频率分别比野生型细胞高约20倍和约110倍。免疫荧光染色显示,这些突变体还表现出异常长且数量众多的细胞质微管束。此外,我们偶尔观察到有丝分裂纺锤体定向异常,完全位于其中一个细胞内。可能是由于纺锤体定向异常而进行核分裂的结果,很大比例的cdp1细胞积累了多个细胞核。虽然cdp1突变体细胞对破坏微管的化合物噻苯咪唑高度敏感,但相对于野生型细胞,它们对密切相关的化合物苯菌灵表现出更高的抗性。综上所述,这些结果表明Cdp1p在控制细胞内微管蛋白动力学中起作用,并且可能直接与微管或微管蛋白相互作用。