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首页> 外文期刊>Molecular and Cellular Biology >Regulation of chromosome segregation by Glc8p, a structural homolog of mammalian inhibitor 2 that functions as both an activator and an inhibitor of yeast protein phosphatase 1.
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Regulation of chromosome segregation by Glc8p, a structural homolog of mammalian inhibitor 2 that functions as both an activator and an inhibitor of yeast protein phosphatase 1.

机译：Glc8p调节染色体分离，Glc8p是哺乳动物抑制剂2的结构同源物，它既是酵母蛋白磷酸酶1的激活剂又是抑制剂。

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The Ipl1 protein kinase is essential for proper chromosome segregation and cell viability in the budding yeast Saccharomyces cerevisiae. We have previously shown that the temperature-sensitive growth phenotype of conditional ipl1-1ts mutants can be suppressed by a partial loss-of-function mutation in the GLC7 gene, which encodes the catalytic subunit (PP1C) of protein phosphatase 1, thus suggesting that this enzyme acts in opposition to the Ipl1 protein kinase in regulating yeast chromosome segregation. We report here that the Glc8 protein, which is related in primary sequence to mammalian inhibitor 2, also participates in this regulation. Like inhibitor 2, the Glc8 protein is heat stable, exhibits anomalous electrophoretic mobility, and functions in vitro as an inhibitor of yeast as well as rabbit skeletal muscle PP1C. Interestingly, overexpression as well as deletion of the GLC8 gene results in a partial suppression of the temperature-sensitive growth phenotype of ipl1ts mutants and also moderately reduces the amount of protein phosphatase 1 activity which is assayable in crude yeast lysates. In addition, the chromosome missegregation phenotype caused by an increase in the dosage of GLC7 is totally suppressed by the glc8-delta 101::LEU2 deletion mutation. These findings together suggest that the Glc8 protein is involved in vivo in the activation of PP1C and that when the Glc8 protein is overproduced, it may also inhibit PP1C function. Furthermore, site-directed mutagenesis studies of GLC8 suggest that Thr-118 of the Glc8 protein, which is equivalent to Thr-72 of inhibitor 2, may play a central role in the ability of this protein to activate and/or inhibit PP1C in vivo.
机译：Ipl1蛋白激酶对于发芽酵母酿酒酵母中正确的染色体分离和细胞活力至关重要。我们以前已经表明，条件性ipl1-1ts突变体的温度敏感型生长表型可以被GLC7基因中的部分功能丧失突变所抑制，该突变编码蛋白磷酸酶1的催化亚基（PP1C），因此表明该酶在调节酵母染色体分离中与Ipl1蛋白激酶相反。我们在这里报告，Glc8蛋白，其在一级序列上与哺乳动物抑制剂2有关，也参与了这一调节。像抑制剂2一样，Glc8蛋白是热稳定的，表现出异常的电泳迁移率，并且在体外起酵母和兔骨骼肌PP1C抑制剂的作用。有趣的是，GLC8基因的过度表达和缺失会导致ipl1ts突变体对温度敏感的生长表型的部分抑制，并且还会适度降低可在粗酵母裂解物中测定的蛋白磷酸酶1的活性。此外，glc8-delta 101 :: LEU2缺失突变完全抑制了由GLC7剂量增加引起的染色体错聚表型。这些发现共同表明，Glc8蛋白在体内参与PP1C的活化，并且当Glc8蛋白过量产生时，它也可能抑制PP1C的功能。此外，对GLC8的定点诱变研究表明，与抑制剂2的Thr-72等效的Glc8蛋白的Thr-118可能在该蛋白体内激活和/或抑制PP1C的能力中起着核心作用。

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