Frederick D L, Tatchell K
Department of Genetics, North Carolina State University, Raleigh 27695, USA.
Mol Cell Biol. 1996 Jun;16(6):2922-31. doi: 10.1128/MCB.16.6.2922.
The GLC7 gene of Saccharomyces cerevisiae encodes the catalytic subunit of type 1 protein phosphatase (PP1) and is essential for cell growth. We have isolated a previously uncharacterized gene, REG2, on the basis of its ability to interact with Glc7p in the two-hybrid system. Reg2p interacts with Glc7p in vivo, and epitope-tagged derivatives of Reg2p and Glc7p coimmunoprecipitate from cell extracts. The predicted protein product of the REG2 gene is similar to Reg1p, a protein believed to direct PP1 activity in the glucose repression pathway. Mutants with a deletion of reg1 display a mild slow-growth defect, while reg2 mutants exhibit a wild-type phenotype. However, mutants with deletions of both reg1 and reg2 exhibit a severe growth defect. Overexpression of REG2 complements the slow-growth defect of a reg1 mutant but does not complement defects in glycogen accumulation or glucose repression, two traits also associated with a reg1 deletion. These results indicate that REG1 has a unique role in the glucose repression pathway but acts together with REG2 to regulate some as yet uncharacterized function important for growth. The growth defect of a reg1 reg2 double mutant is alleviated by a loss-of-function mutation in the SNF1-encoded protein kinase. The snf1 mutation also suppresses the glucose repression defects of reg1. Together, our data are consistent with a model in which Reg1p and Reg2p control the activity of PP1 toward substrates that are phosphorylated by the Snf1p kinase.
酿酒酵母的GLC7基因编码1型蛋白磷酸酶(PP1)的催化亚基,对细胞生长至关重要。我们基于其在双杂交系统中与Glc7p相互作用的能力,分离出了一个先前未被表征的基因REG2。Reg2p在体内与Glc7p相互作用,并且Reg2p和Glc7p的表位标记衍生物可从细胞提取物中共免疫沉淀。REG2基因的预测蛋白产物与Reg1p相似,Reg1p是一种被认为在葡萄糖阻遏途径中指导PP1活性的蛋白质。缺失reg1的突变体表现出轻微的生长缓慢缺陷,而reg2突变体表现出野生型表型。然而,同时缺失reg1和reg2的突变体表现出严重的生长缺陷。REG2的过表达弥补了reg1突变体的生长缓慢缺陷,但不能弥补糖原积累或葡萄糖阻遏方面的缺陷,这两个特征也与reg1缺失有关。这些结果表明Reg1在葡萄糖阻遏途径中具有独特作用,但与REG2共同作用以调节某些对生长很重要但尚未表征的功能。SNF1编码的蛋白激酶功能丧失突变可缓解reg1 reg2双突变体的生长缺陷。snf1突变也抑制了reg1的葡萄糖阻遏缺陷。总之,我们的数据与一个模型一致,即Reg1p和Reg2p控制PP1对被Snf1p激酶磷酸化的底物的活性。