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首页> 外文期刊>Molecular and Cellular Biology >The Phosphotyrosyl Phosphatase Activator, Ncs1p (Rrd1p), Functions with Cla4p To Regulate the G2/M Transition inSaccharomyces cerevisiae
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The Phosphotyrosyl Phosphatase Activator, Ncs1p (Rrd1p), Functions with Cla4p To Regulate the G2/M Transition inSaccharomyces cerevisiae

机译：磷酸酪氨酰磷酸酶激活剂Ncs1p（Rrd1p）与Cla4p一起调控酿酒酵母中的G2 / M过渡

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The Saccharomyces cerevisiae p21-activated kinases, Ste20p and Cla4p, have individual functions but appear to share an essential function(s) as well because a strain lacking both kinases is inviable. To learn more about the shared function, we sought new mutations that were lethal in the absence of CLA4. This approach led to the identification of at least 10 complementation groups designated NCS (need CLA4 to survive). As for ste20 cla4-75 mutants, most ncs cla4-75double mutants were defective for septin localization during budding. One group, NCS1/RRD1 (YIL153w), did not confer this defect, however, and we investigated its function further.ncs1Δ cla4Δ cells arrested with elongated buds and short mitotic spindles. The morphological defects and lethality were suppressed by mutations that abrogate the cell cycle morphogenetic checkpoint, CDC28Y19F or swe1Δ. The connection to the cell cycle may be direct, as we detected a Cla4p-Cdc28p complex. NCS1 encodes a protein with significant similarity to a mammalian phosphotyrosyl phosphatase activator (PTPA) regulatory subunit for type 2A protein phosphatases (PP2As). Genetic and biochemical evidence suggested that the phosphatase Sit4p is a target for Ncs1p. First, CLA4 andSIT4 were synthetically lethal. Second, Ncs1p and its yeast paralog, Noh1p (Rrd2p), bound to the catalytic domain of Sit4p in vitro, and Ncs1p could be immunoprecipitated with Sit4p but not with another PP2A (Pph21p) from yeast cell extracts. Strains lacking bothNCS1 and NOH1 were inviable and arrested as unbudded cells, implying that PTPA function is required for proper G1 progression.
机译：酿酒酵母（Saccharomyces cerevisiae）p21激活的激酶Ste20p和Cla4p具有单独的功能，但似乎也具有基本功能，因为缺乏这两种激酶的菌株是不可行的。要了解有关共享功能的更多信息，我们寻求了在没有 CLA4 的情况下具有致命性的新突变。这种方法导致鉴定出至少10个称为 NCS 的互补组（需要 CLA4 才能存活）。至于 ste20 cla4-75 突变体，大多数 ncs cla4-75 double突变体在发芽过程中对Septin定位都是有缺陷的。一组 NCS1 / RRD1 （ YIL153w ）没有造成这种缺陷，但是我们进一步研究了其功能。ncs1Δcla4 Δ细胞具长芽和短有丝分裂纺锤体被捕。消除细胞周期形态发生检查点 CDC28Y19F 或 swe1 Δ的突变抑制了形态缺陷和致死性。由于我们检测到Cla4p-Cdc28p复合体，因此与细胞周期的联系可能是直接的。 NCS1 编码的蛋白与2A型蛋白磷酸酶（PP2As）的哺乳动物磷酸酪氨酰磷酸酶激活剂（PTPA）调节亚基极为相似。遗传和生化证据表明，磷酸酶Sit4p是Ncs1p的靶标。首先， CLA4 和 SIT4 具有综合致命性。其次，Ncs1p及其酵母类似物Noh1p（Rrd2p）在体外与Sit4p的催化结构域结合，并且Ncs1p可以用Sit4p免疫沉淀，而不能用来自酵母细胞提取物的另一PP2A（Pph21p）进行免疫沉淀。缺乏 NCS1 和 NOH1 的菌株是不能存活的，并且被捕为未预算的细胞，这表明PTPA功能是G 1 正常发育所必需的。

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