Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35487, United States.
Biochem Biophys Res Commun. 2010 Sep 3;399(4):665-9. doi: 10.1016/j.bbrc.2010.07.139. Epub 2010 Aug 4.
In the fission yeast, Schizosaccharomyces pombe, cyclic AMP (cAMP)-dependent protein kinase (PKA) is not essential for viability under normal culturing conditions, making this organism attractive for investigating mechanisms of PKA regulation. Here we show that S. pombe cells carrying a deletion in the adenylate cyclase gene, cyr1, express markedly higher levels of the PKA catalytic subunit, Pka1, than wild type cells. Significantly, in cyr1Delta cells, but not wild type cells, a substantial proportion of Pka1 protein is hyperphosphorylated. Pka1 hyperphosphorylation is strongly induced in cyr1Delta cells, and to varying degrees in wild type cells, by both glucose starvation and stationary phase stresses, which are associated with reduced cAMP-dependent PKA activity, and by KCl stress, the cellular adaptation to which is dependent on PKA activity. Interestingly, hyperphosphorylation of Pka1 was not detected in either cyr1(+) or cyr1Delta S. pombe strains carrying a deletion in the PKA regulatory subunit gene, cgs1, under any of the tested conditions. Our results demonstrate the existence of a cAMP-independent mechanism of PKA catalytic subunit phosphorylation, which we propose could serve as a mechanism for inducing or maintaining specific PKA functions under conditions in which its cAMP-dependent activity is downregulated.
在裂殖酵母(Schizosaccharomyces pombe)中,环腺苷酸(cyclic AMP,cAMP)依赖性蛋白激酶(protein kinase A,PKA)在正常培养条件下并非生存所必需,这使得该生物成为研究 PKA 调节机制的理想对象。本文研究表明,携带环腺苷酸合成酶基因(cyr1)缺失的裂殖酵母细胞表达的 PKA 催化亚基 Pka1 的水平明显高于野生型细胞。值得注意的是,在 cyr1Δ细胞中,而非野生型细胞中,相当一部分 Pka1 蛋白发生过度磷酸化。cyr1Δ细胞中 Pka1 的过度磷酸化可被葡萄糖饥饿和静止期应激强烈诱导,并在一定程度上被 KCl 应激诱导,这些应激条件均与 cAMP 依赖性 PKA 活性降低有关,而细胞对 KCl 应激的适应则依赖于 PKA 活性。有趣的是,在任何测试条件下,cyr1(+)或 cyr1Δ裂殖酵母菌株中,当 PKA 调节亚基基因(cgs1)缺失时,均未检测到 Pka1 的过度磷酸化。我们的研究结果表明,存在一种 cAMP 非依赖性的 PKA 催化亚基磷酸化机制,我们推测这种机制可作为在 cAMP 依赖性活性下调的条件下诱导或维持特定 PKA 功能的机制。