Hisamoto N, Frederick D L, Sugimoto K, Tatchell K, Matsumoto K
Department of Molecular Biology, Faculty of Science, Nagoya University, Japan.
Mol Cell Biol. 1995 Jul;15(7):3767-76. doi: 10.1128/MCB.15.7.3767.
The Saccharomyces cerevisiae GLC7 gene encodes the catalytic subunit of type 1 protein phosphatase (PP1) and is required for cell growth. A cold-sensitive glc7 mutant (glc7Y170) arrests in G2/M but remains viable at the restrictive temperature. In an effort to identify additional gene products that function in concert with PP1 to regulate growth, we isolated a mutation (gpp1) that exacerbated the growth phenotype of the glc7Y170 mutation, resulting in rapid death of the double mutant at the nonpermissive temperature. We identified an additional gene, EGP1, as an extra-copy suppressor of the glc7Y170 gpp1-1 double mutant. The nucleotide sequence of EGP1 predicts a leucine-rich repeat protein that is similar to Sds22, a protein from the fission yeast Schizosaccharomyces pombe that positively modulates PP1. EGP1 is essential for cell growth but becomes dispensable upon overexpression of the GLC7 gene. Egp1 and PP1 directly interact, as assayed by coimmunoprecipitation. These results suggest that Egp1 functions as a positive modulator of PP1 in the growth control of S. cerevisiae.
酿酒酵母的GLC7基因编码1型蛋白磷酸酶(PP1)的催化亚基,是细胞生长所必需的。一个冷敏感的glc7突变体(glc7Y170)在G2/M期停滞,但在限制温度下仍可存活。为了鉴定与PP1协同作用以调节生长的其他基因产物,我们分离出一个突变(gpp1),它加剧了glc7Y170突变的生长表型,导致双突变体在非允许温度下迅速死亡。我们鉴定出另一个基因EGP1,作为glc7Y170 gpp1-1双突变体的额外拷贝抑制子。EGP1的核苷酸序列预测其为富含亮氨酸重复序列的蛋白,与粟酒裂殖酵母中的Sds22蛋白相似,后者对PP1起正向调节作用。EGP1对细胞生长至关重要,但在GLC7基因过表达时变得不再必需。通过免疫共沉淀分析表明,Egp1和PP1直接相互作用。这些结果表明,Egp1在酿酒酵母的生长控制中作为PP1的正向调节因子发挥作用。