The dual-specificity protein phosphatase Yvh1p regulates sporulation, growth, and glycogen accumulation independently of catalytic activity in Saccharomyces cerevisiae via the cyclic AMP-dependent protein kinase cascade
The dual-specificity protein phosphatase Yvh1p regulates sporulation, growth, and glycogen accumulation independently of catalytic activity in Saccharomyces cerevisiae via the cyclic AMP-dependent protein kinase cascade
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DOI：
10.1128/jb.182.12.3517-3528.2000
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发表时间：
2000-06-01
影响因子：
3.2
通讯作者：
Cooper, TG
中科院分区：
文献类型：
--
作者：
Beeser, AE;Cooper, TG
Yvh1p是一种由氮饥饿诱导的双特异性蛋白磷酸酶，调节细胞生长以及产孢量的启动和完成。我证明了yvh1中断突变体也不能在固定相积累糖原。Yvh1(C117S)的一个催化失活变异体和一个只编码Yvh1p C端159个氨基酸的DNA片段(完全缺乏磷酸酶结构域)补充了所有三种表型以及野生型等位基因；只编码C端74个氨基酸的片段没有互补作用。这些观察结果表明，我们测量的Yvh1p功能不需要磷酸酶活性。降低内源性循环AR；IP(CAMP)水平的突变部分抑制了产孢子和糖原积累缺陷。此外，在yvh1中断突变体中，由DRR2启动子片段支持的报告基因表达减少，该片段包含两个已知对cAMP-蛋白激酶A做出反应的应激反应元件。因此，我们的结果识别了三个细胞过程，这三个过程都需要Yvh1p并对cAMP的变化做出反应，这使得我们认为Yvh1p可能是cAMP依赖的蛋白激酶级联调控的参与者和/或贡献者。事实上，降低cAMP水平可以减轻对Yvh1p功能的需求，这一事实支持这一观点。
Yvh1p, a dual-specific protein phosphatase induced specifically by nitrogen starvation, regulates cell growth as well as initiation and completion of sporulation. me demonstrate that yvh1 disruption mutants are also unable to accumulate glycogen in stationary phase. A catalytically inactive variant of yvh1 (C117S) and a DNA fragment encoding only the Yvh1p C-terminal 159 amino acids (which completely lacks the phosphatase domain) complement all three phenotypes as well as the wild-type allele; no complementation occurs with a fragment encoding only the C-terminal 74 amino acids. These observations argue that phosphatase activity is not required for the Yvh1p functions we measured. Mutations which decrease endogenous cyclic AR;IP (cAMP) levels partially suppress the sporulation and glycogen accumulation defects. In addition, reporter gene expression supported by a DRR2 promoter fragment, containing two stress response elements known to respond to cAMP-protein kinase A, decreases in a yvh1 disruption mutant. Therefore, our results identify three cellular processes that both require Yvh1p and respond to alterations in cAMP, and they lead us to suggest that Yvh1p may be a participant in and/or a contributor to regulation of the cAMP-dependent protein kinase cascade. The fact that decreasing the levels of cAMP alleviates the need for Yvh1p function supports this suggestion.