Matsuzawa Tomohiko, Fujita Yasuko, Tohda Hideki, Takegawa Kaoru
Department of Bioscience & Biotechnology, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
Eukaryot Cell. 2012 Feb;11(2):159-67. doi: 10.1128/EC.05268-11. Epub 2011 Dec 2.
The function of two fission yeast genes, SPCC74.03c/ssp2(+) and SPAC23H4.02/ppk9(+), encoding an Snf1-like protein kinase were investigated. Deletion of ssp2(+) caused a partial defect in glucose derepression of inv1(+), fbp1(+), and gld1(+) and in assimilation of sucrose and glycerol, while a mutation in ppk9(+) had no apparent effect. Scr1, a transcription factor involved in glucose repression, localized to the nucleus under glucose-rich conditions and to the cytoplasm during glucose starvation in wild-type cells. In contrast, in the ssp2Δ mutant, Scr1 localized to the nucleus in cells grown in glucose-rich medium as well as in glucose-starved cells. Immunoblot analysis showed that Ssp2 is required for the phosphorylation of Scr1 upon glucose deprivation. Mutation of five putative Ssp2 recognition sites in Scr1 prevented glucose derepression of invertase in glucose-starved cells. These results indicate that Ssp2 regulates phosphorylation and subcellular localization of Scr1 in response to glucose.
对编码一种Snf1样蛋白激酶的两个裂殖酵母基因SPCC74.03c/ssp2(+)和SPAC23H4.02/ppk9(+)的功能进行了研究。缺失ssp2(+)会导致inv1(+)、fbp1(+)和gld1(+)的葡萄糖去阻遏以及蔗糖和甘油同化方面出现部分缺陷,而ppk9(+)的突变则无明显影响。Scr1是一种参与葡萄糖阻遏的转录因子,在野生型细胞中,在富含葡萄糖的条件下定位于细胞核,在葡萄糖饥饿期间定位于细胞质。相反,在ssp2Δ突变体中,Scr1在富含葡萄糖的培养基中生长的细胞以及葡萄糖饥饿的细胞中均定位于细胞核。免疫印迹分析表明,葡萄糖剥夺时,Ssp2是Scr1磷酸化所必需的。Scr1中五个假定的Ssp2识别位点的突变阻止了葡萄糖饥饿细胞中转化酶的葡萄糖去阻遏。这些结果表明,Ssp2响应葡萄糖调节Scr1的磷酸化和亚细胞定位。