Xu B E, Skowronek K R, Kurjan J
Department of Microbiology and Molecular Genetics and the Vermont Cancer Center, University of Vermont, College of Medicine, Burlington, Vermont 05405-0068, USA.
Genetics. 2001 Dec;159(4):1559-71. doi: 10.1093/genetics/159.4.1559.
The Saccharomyces cerevisiae RGS protein Sst2p is involved in desensitization to pheromone and acts as a GTPase-activating protein for the Galpha subunit Gpa1p. Other results indicate that Sst2p acts through Mpt5p and that this action occurs downstream of Fus3p and through Cln3p/Cdc28p. Our results indicate that the interaction of Sst2p with Mpt5p requires the N-terminal MPI (Mpt5p-interacting) domain of Sst2p and is independent of the C-terminal RGS domain. Overexpression of the MPI domain results in an Mpt5p-dependent increase in recovery from pheromone arrest. Overexpression of either intact Sst2p or the MPI domain leads to partial suppression of a gpa1 growth defect, and this suppression is dependent on Mpt5p, indicating that MPI function occurs downstream of Gpa1p and through Mpt5p. Combination of an mpt5 mutation with the GPA1(G302S) mutation, which uncouples Gpa1p from Sst2p, results in pheromone supersensitivity similar to the sst2 mutant, and promotion of recovery by overexpression of Sst2p is dependent on both Mpt5p and the Gpa1p interaction. These results indicate that Sst2p is a bifunctional protein and that the MPI domain acts through Mpt5p independently of the RGS domain. RGS family members from other fungi contain N-terminal domains with sequence similarity to the Sst2p MPI domain, suggesting that MPI function may be conserved.
酿酒酵母的RGS蛋白Sst2p参与对信息素的脱敏作用,并作为Gα亚基Gpa1p的GTP酶激活蛋白。其他结果表明,Sst2p通过Mpt5p发挥作用,且这种作用发生在Fus3p的下游并通过Cln3p/Cdc28p。我们的结果表明,Sst2p与Mpt5p的相互作用需要Sst2p的N端MPI(与Mpt5p相互作用)结构域,且独立于C端RGS结构域。MPI结构域的过表达导致在信息素诱导的停滞恢复中依赖Mpt5p的增加。完整的Sst2p或MPI结构域的过表达导致gpa1生长缺陷的部分抑制,且这种抑制依赖于Mpt5p,表明MPI功能发生在Gpa1p的下游并通过Mpt5p。mpt5突变与GPA1(G302S)突变(使Gpa1p与Sst2p解偶联)的组合导致类似于sst2突变体的信息素超敏反应,并且Sst2p过表达促进恢复依赖于Mpt5p和Gpa1p相互作用。这些结果表明,Sst2p是一种双功能蛋白,且MPI结构域独立于RGS结构域通过Mpt5p发挥作用。来自其他真菌的RGS家族成员含有与Sst2p MPI结构域具有序列相似性的N端结构域,表明MPI功能可能是保守的。