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Cross regulation of four GATA factors that control nitrogen catabolic gene expression in Saccharomyces cerevisiae.

机译:调控啤酒酵母中氮分解代谢基因表达的四个GATA因子的交叉调控。

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摘要

Nitrogen catabolic gene expression in Saccharomyces cerevisiae has been reported to be regulated by three GATA family proteins, the positive regulators Gln3p and Gat1p/Nil1p and the negative regulator Dal80p/Uga43p. We show here that a fourth member of the yeast GATA family, the Dal80p homolog Deh1p, also negatively regulates expression of some, but not all, nitrogen catabolic genes, i.e., GAP1, DAL80, and UGA4 expression increases in a deh1 delta mutant. Consistent with Deh1p regulation of these genes is the observation that Deh1p forms specific DNA-protein complexes with GATAA-containing UGA4 and GAP1 promoter fragments in electrophoretic mobility shift assays. Deh1p function is demonstrable, however, only when a repressive nitrogen source such as glutamine is present; deh1 delta mutants exhibit no detectable phenotype with a poor nitrogen source such as proline. Our experiments also demonstrate that GATA factor gene expression is highly regulated by the GATA factors themselves in an interdependent manner. DAL80 expression is Gln3p and Gat1p dependent and Dal80p regulated. Moreover, Gln3p and Dal80p bind to DAL80 promoter fragments. In turn, GAT1 expression is Gln3p dependent and Dal80p regulated but is not autogenously regulated like DAL80. DEH1 expression is largely Gln3p independent, modestly Gat1p dependent, and most highly regulated by Dal80p. Paradoxically, the high-level DEH1 expression observed in a dal80::hisG disruption mutant is highly sensitive to nitrogen catabolite repression.
机译:据报道,酿酒酵母中的氮分解代谢基因表达受到三种GATA家族蛋白的调节,即正调节剂Gln3p和Gat1p / Nil1p和负调节剂Dal80p / Uga43p。我们在这里显示了酵母GATA家族的第四个成员Dal80p同源物Deh1p也负调节一些但不是全部的氮分解代谢基因(即GAP1,DAL80和UGA4的表达)在deh1 delta突变体中的表达增加。与这些基因的Deh1p调控一致的是,在电泳迁移率迁移分析中,Deh1p与含GATAA的UGA4和GAP1启动子片段形成了特定的DNA-蛋白质复合物。然而,只有当存在抑制性氮源(如谷氨酰胺)时,Deh1p的功能才能证明。 deh1 delta突变体没有显示出可检测的表型,氮源较弱,如脯氨酸。我们的实验还证明,GATA因子基因的表达受到GATA因子本身的高度调控,相互依赖。 DAL80表达受Gln3p和Gat1p依赖,且受Dal80p调控。此外,Gln3p和Dal80p与DAL80启动子片段结合。反过来,GAT1表达是Gln3p依赖性和Dal80p调节的,但不像DAL80那样自发地调节。 DEH1表达在很大程度上与Gln3p无关,与Gat1p无关,并且受Dal80p高度调节。矛盾的是,在dal80 :: hisG破坏突变体中观察到的高水平DEH1表达对氮分解代谢物阻遏高度敏感。

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