Yang X, Jiang R, Carlson M
Department of Genetics, College of Physicians and Surgeons of Columbia University, New York, NY 10032.
EMBO J. 1994 Dec 15;13(24):5878-86. doi: 10.1002/j.1460-2075.1994.tb06933.x.
The SNF1 protein kinase is required for the regulatory response to glucose starvation in Saccharomyces cerevisiae. SNF1 is a protein serine/threonine kinase that has been widely conserved in both plants and mammals. Previously, we identified SIP1 and SIP2 as proteins that interact with SNF1 in vivo by the two-hybrid system. We have cloned the SIP2 gene and the encoded protein is homologous to SIP1 and to GAL83, which affects glucose repression of the GAL genes. We show that SIP2 and GAL83, like SIP1, co-immunoprecipitate with SNF1 and are phosphorylated in vitro. An 80 amino acid sequence, designated the ASC domain, is highly conserved at the C-termini of all three proteins. We show that this small domain can mediate protein-protein interaction with the SNF1 kinase complex. Thus, SIP1, SIP2 and GAL83 define a family of homologous proteins that are tightly associated with the SNF1 kinase, probably in alternative forms of the complex. Genetic evidence suggests that the three proteins have distinct, but related, functions in the SNF1 pathway, and deletion of GAL83 dramatically reduces SNF1 activity in immune complex assays. We propose that SIP1, SIP2 and GAL83 act as adaptors that promote the activity of SNF1 towards specific targets.
SNF1蛋白激酶是酿酒酵母对葡萄糖饥饿进行调节反应所必需的。SNF1是一种蛋白丝氨酸/苏氨酸激酶,在植物和哺乳动物中都广泛保守。此前,我们通过双杂交系统鉴定出SIP1和SIP2是在体内与SNF1相互作用的蛋白。我们克隆了SIP2基因,其编码的蛋白与SIP1以及影响GAL基因葡萄糖阻遏的GAL83同源。我们发现,SIP2和GAL83与SIP1一样,能与SNF1进行共免疫沉淀,并且在体外被磷酸化。一个80个氨基酸的序列,称为ASC结构域,在这三种蛋白的C末端高度保守。我们表明,这个小结构域能介导与SNF1激酶复合物的蛋白-蛋白相互作用。因此,SIP1、SIP2和GAL83定义了一个同源蛋白家族,它们与SNF1激酶紧密相关,可能以复合物的不同形式存在。遗传证据表明,这三种蛋白在SNF1途径中具有不同但相关的功能,并且在免疫复合物分析中,GAL83的缺失会显著降低SNF1活性。我们提出,SIP1、SIP2和GAL83作为衔接蛋白,促进SNF1对特定靶标的活性。