Ryu S, Garges S
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
J Biol Chem. 1994 Feb 18;269(7):4767-72.
The ptsH operon of Escherichia coli is controlled by two promoters P0 and P1, each of which is regulated by cyclic AMP receptor protein (CRP) complexed with cAMP (CRP.cAMP). We have studied the in vitro as well as in vivo transcriptional regulation of these two promoters. Each promoter exhibits a switching mechanism in vitro, where, depending upon the presence or absence of CRP.cAMP, transcription is initiated from different start sites termed a and b. P0 (P0a) is affected by supercoiling: when the template is linear, transcription initiation is switched to a site 3 base pairs upstream (P0b) and becomes more CRP.cAMP dependent. Transcription from the P1 promoter (P1a) switches initiation sites to 7 base pairs downstream (P1b) in the presence of CRP.cAMP. Most transcription in vivo was from P1a, and P0b could not be detected in vivo. Glucose has independent positive effects on pts expression in vivo. The results indicate that the two different regulatory mechanisms (one through CRP.cAMP, the other through glucose) are working together for fine control of pts expression.
大肠杆菌的ptsH操纵子由两个启动子P0和P1控制,每个启动子都由与环磷酸腺苷(cAMP)复合的环磷酸腺苷受体蛋白(CRP)(CRP.cAMP)调控。我们研究了这两个启动子的体外和体内转录调控。每个启动子在体外都表现出一种切换机制,根据CRP.cAMP的存在与否,转录从不同的起始位点开始,分别称为a和b。P0(P0a)受超螺旋影响:当模板为线性时,转录起始切换到上游3个碱基对的位点(P0b),并且变得更依赖CRP.cAMP。在存在CRP.cAMP的情况下,来自P1启动子(P1a)的转录起始位点切换到下游7个碱基对(P1b)。体内的大多数转录来自P1a,并且在体内无法检测到P0b。葡萄糖在体内对pts表达具有独立的正向作用。结果表明,两种不同的调控机制(一种通过CRP.cAMP,另一种通过葡萄糖)共同作用以精细控制pts表达。