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DBF2蛋白激酶与细胞周期调控的MOB1蛋白结合并通过其发挥作用。

DBF2 protein kinase binds to and acts through the cell cycle-regulated MOB1 protein.

作者信息

Komarnitsky S I, Chiang Y C, Luca F C, Chen J, Toyn J H, Winey M, Johnston L H, Denis C L

机构信息

Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham 03824, USA.

出版信息

Mol Cell Biol. 1998 Apr;18(4):2100-7. doi: 10.1128/MCB.18.4.2100.

Abstract

The DBF2 gene of the budding yeast Saccharomyces cerevisiae encodes a cell cycle-regulated protein kinase that plays an important role in the telophase/G1 transition. As a component of the multisubunit CCR4 transcriptional complex, DBF2 is also involved in the regulation of gene expression. We have found that MOB1, an essential protein required for a late mitotic event in the cell cycle, genetically and physically interacts with DBF2. DBF2 binds MOB1 in vivo and can bind it in vitro in the absence of other yeast proteins. We found that the expression of MOB1 is also cell cycle regulated, its expression peaking slightly before that of DBF2 at the G2/M boundary. While overexpression of DBF2 suppressed phenotypes associated with mob1 temperature-sensitive alleles, it could not suppress a mob1 deletion. In contrast, overexpression of MOB1 suppressed phenotypes associated with a dbf2-deleted strain and suppressed the lethality associated with a dbf2 dbf20 double deletion. A mob1 temperature-sensitive allele with a dbf2 disruption was also found to be synthetically lethal. These results are consistent with DBF2 acting through MOB1 and aiding in its function. Moreover, the ability of temperature-sensitive mutated versions of the MOB1 protein to interact with DBF2 was severely reduced, confirming that binding of DBF2 to MOB1 is required for a late mitotic event. While MOB1 and DBF2 were found to be capable of physically associating in a complex that did not include CCR4, MOB1 did interact with other components of the CCR4 transcriptional complex. We discuss models concerning the role of DBF2 and MOB1 in controlling the telophase/G1 transition.

摘要

出芽酵母酿酒酵母的DBF2基因编码一种细胞周期调控蛋白激酶,该激酶在末期/G1期转换中起重要作用。作为多亚基CCR4转录复合体的一个组成部分,DBF2也参与基因表达的调控。我们发现,MOB1是细胞周期中一个有丝分裂后期事件所必需的一种重要蛋白质,它在遗传和物理上与DBF2相互作用。DBF2在体内与MOB1结合,并且在没有其他酵母蛋白的情况下也能在体外与它结合。我们发现MOB1的表达也是细胞周期调控的,其表达在G2/M边界处略早于DBF2达到峰值。虽然DBF2的过表达抑制了与mob1温度敏感等位基因相关的表型,但它不能抑制mob1缺失。相反,MOB1的过表达抑制了与dbf2缺失菌株相关的表型,并抑制了与dbf2 dbf20双缺失相关的致死性。还发现一个带有dbf2破坏的mob1温度敏感等位基因是合成致死的。这些结果与DBF2通过MOB1发挥作用并辅助其功能是一致的。此外,MOB1蛋白的温度敏感突变体与DBF2相互作用的能力严重降低,这证实了DBF2与MOB1的结合是有丝分裂后期事件所必需的。虽然发现MOB1和DBF2能够在不包括CCR4的复合体中进行物理结合,但MOB1确实与CCR4转录复合体的其他组分相互作用。我们讨论了关于DBF2和MOB1在控制末期/G1期转换中作用的模型。

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