Phospholipase C interacts with Sgd1p and is required for expression of GPD1 and osmoresistance in Saccharomyces cerevisiae.

Phospholipase C interacts with Sgd1p and is required for expression of GPD1 and osmoresistance in Saccharomyces cerevisiae.
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磷脂酶 C 与 Sgd1p 相互作用,是酿酒酵母中 GPD1 表达和渗透阻力所必需的。

DOI:
10.1007/s00438-002-0647-8
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发表时间:
2002
期刊:
Molecular genetics and genomics : MGG.
影响因子:
--
通讯作者:
Vancura,A
Vancura,A
中科院分区:
--
文献类型:
--
作者:
Lin,H;Nguyen,P;Vancura,A

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酿酒酵母PLC 1基因编码哺乳动物磷酸肌醇特异性磷脂酶C的δ亚型同源物。缺失PLC 1的细胞(plc 1 Δ)是活的,但显示出几种表型,包括渗透、温度和诺考达唑敏感性。我们已经使用了双杂交筛选,以确定Plc 1 p相互作用的蛋白质。发现的相互作用蛋白之一是Sgd 1 p,这是一种最近发现的必需核蛋白。SGD 1基因最初是通过互补一个对水稻胁迫敏感的突变体而克隆的。通过亲和层析证实了Plc 1 p-Sgd 1 p相互作用。SGD 1与PLC 1和HOG 1(其编码一种对有丝分裂原活化蛋白激酶敏感的蛋白激酶)都有遗传相互作用。Sgd 1 p的过表达抑制了携带plc 1 - 4等位基因的细胞的温度敏感性,双突变株plc 1 Δ sgd 1 - 1显示出增强的温度和诺考达唑敏感性。plc 1 Δ hog 1 Δ菌株显示出增加的甘油敏感性,并且在甘油合成和GPD 1(其编码参与甘油生物合成的酶甘油3-磷酸脱氢酶)的表达中具有合成缺陷,这表明Plc 1 p和Hog 1 p在独立的途径中发挥作用。hog 1 Δ sgd 1 - 1双突变体的荧光敏感性比单突变体的荧光敏感性高。三重质粒plc 1 Δ hog 1 Δ sgd 1 - 1是无活力的,而plc 1 Δ hog 1 Δ sgd 1 - 2菌株生长极其缓慢,并且比plc 1 Δ hog 1 Δ orhog 1 Δ sgd 1 - 2菌株更敏感。这些结果是一致的模型,其中Plc 1 p和Hog 1 p的功能在平行的途径影响β调节,和信号从这两个途径收敛,至少部分地,在Sgd 1 p。
TheSaccharomyces cerevisiae PLC1gene encodes a homolog of the δ isoform of mammalian phosphoinositide-specific phospholipase C. Cells deleted forPLC1(plc1Δ) are viable, but display several phenotypes, including osmotic, temperature, and nocodazole sensitivity. We have used a two-hybrid screen to identify Plc1p-interacting proteins. One of the interacting proteins found was Sgd1p, a recently identified, essential, nuclear protein. TheSGD1gene was originally cloned by complementation of an osmostress-sensitive mutant. The Plc1p-Sgd1p interaction was confirmed biochemically by affinity chromatography.SGD1interacts genetically with bothPLC1andHOG1(which encodes an osmosensing mitogen-activated protein kinase). Overexpression of Sgd1p suppresses the temperature sensitivity of cells bearing theplc1-4allele, and the double mutant strainplc1Δsgd1-1displays enhanced temperature and nocodazole sensitivity. Theplc1Δhog1Δ strain displays increased osmosensitivity, and has a synthetic defect in glycerol synthesis and the expression ofGPD1(which encodes the enzyme glycerol 3-phosphate dehydrogenase that is involved in glycerol biosynthesis), suggesting that Plc1p and Hog1p function in independent pathways. Thehog1Δsgd1-1double mutant displays enhanced osmosensitivity relative to that of either single mutant. The triple mutantplc1Δhog1Δsgd1-1is inviable, while theplc1Δhog1Δsgd1-2strain grows extremely slowly and is more osmosensitive than theplc1Δhog1Δ orhog1Δsgd1-2strain. These results are consistent with a model in which Plc1p and Hog1p function in parallel pathways affecting osmoregulation, and signals from both these pathways converge, at least partly, on Sgd1p.
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