Coffman J A, Rai R, Cunningham T, Svetlov V, Cooper T G
Department of Microbiology and Immunology, University of Tennessee, Memphis 38163, USA.
Mol Cell Biol. 1996 Mar;16(3):847-58. doi: 10.1128/MCB.16.3.847.
Saccharomyces cerevisiae cells selectively use nitrogen sources in their environment. Nitrogen catabolite repression (NCR) is the basis of this selectivity. Until recently NCR was thought to be accomplished exclusively through the negative regulation of Gln3p function by Ure2p. The demonstration that NCR-sensitive expression of multiple nitrogen-catabolic genes occurs in a gln3 delta ure2 delta dal80::hisG triple mutant indicated that the prevailing view of the nitrogen regulatory circuit was in need of revision; additional components clearly existed. Here we demonstrate that another positive regulator, designated Gat1p, participates in the transcription of NCR-sensitive genes and is able to weakly activate transcription when tethered upstream of a reporter gene devoid of upstream activation sequence elements. Expression of GAT1 is shown to be NCR sensitive, partially Gln3p dependent, and Dal80p regulated. In agreement with this pattern of regulation, we also demonstrate the existence of Gln3p and Dal80p binding sites upstream of GAT1.
酿酒酵母细胞会选择性地利用其环境中的氮源。氮分解代谢物阻遏(NCR)是这种选择性的基础。直到最近,人们还认为NCR完全是通过Ure2p对Gln3p功能的负调控来实现的。多个氮分解代谢基因的NCR敏感型表达在gln3δure2δdal80::hisG三重突变体中出现,这表明氮调节回路的主流观点需要修正;显然还存在其他成分。在这里,我们证明了另一种正调控因子,命名为Gat1p,参与NCR敏感型基因的转录,并且当它与缺乏上游激活序列元件的报告基因上游相连时,能够微弱地激活转录。GAT1的表达显示出对NCR敏感,部分依赖Gln3p,并受Dal80p调控。与这种调控模式一致,我们还证明了在GAT1上游存在Gln3p和Dal80p结合位点。