Hisamoto N, Sugimoto K, Matsumoto K
Department of Molecular Biology, Faculty of Science, Nagoya University, Japan.
Mol Cell Biol. 1994 May;14(5):3158-65. doi: 10.1128/mcb.14.5.3158-3165.1994.
We isolated a mutant carrying a conditional mutation in the GLC7 gene, encoding the catalytic subunit of a type 1 protein phosphatase, by selection of suppressors that restored the growth defect of cdc24 mutants at high temperature and simultaneously conferred cold-sensitive growth. This cold sensitivity for growth is caused by a single mutation (glc7Y-170) at position 170 of the Glc7 protein, resulting in replacement of cysteine with tyrosine. Genetic analysis suggested that the glc7Y-170 allele is associated with a recessive negative phenotype, reducing the activity of Glc7 in the cell. The glc7Y-170 mutant missegregated chromosome III at the permissive temperature, arrested growth as large-budded cells at the restrictive temperature, exhibited a significant increase in the number of nuclei at or in the neck, and had a short spindle. Furthermore, the glc7Y-170 mutant exhibited a high level of CDC28-dependent protein kinase activity when incubated at the restrictive temperature. These findings suggest that the glc7Y-170 mutation is defective in the G2/M phase of the cell cycle. Thus, type 1 protein phosphatase in Saccharomyces cerevisiae is essential for the G2/M transition.
我们通过筛选能够恢复cdc24突变体在高温下生长缺陷并同时赋予冷敏感生长特性的抑制子,分离出了一个在GLC7基因中携带条件突变的突变体,该基因编码1型蛋白磷酸酶的催化亚基。这种对生长的冷敏感性是由Glc7蛋白第170位的单个突变(glc7Y - 170)引起的,导致半胱氨酸被酪氨酸取代。遗传分析表明,glc7Y - 170等位基因与隐性负表型相关,降低了细胞中Glc7的活性。glc7Y - 170突变体在允许温度下会错误分离III号染色体,在限制温度下会以大芽细胞的形式停止生长,在颈部或颈部内的细胞核数量显著增加,并且纺锤体较短。此外,glc7Y - 170突变体在限制温度下孵育时表现出高水平的依赖CDC28的蛋白激酶活性。这些发现表明,glc7Y - 170突变在细胞周期的G2/M期存在缺陷。因此,酿酒酵母中的1型蛋白磷酸酶对于G2/M期转换至关重要。