Gerlach P, Valentin-Hansen P, Bremer E
Department of Biology, University of Konstanz, FRG.
Mol Microbiol. 1990 Mar;4(3):479-88. doi: 10.1111/j.1365-2958.1990.tb00614.x.
The Escherichia coli cytR-encoded repressor protein (CytR) controls the expression of several genes involved in nucleoside and deoxynucleoside uptake and metabolism. The cytR promoter was identified by determining the transcriptional initiation site of the cytR gene. A chromosomal cytR-lacZ+ operon fusion was isolated and used to study the regulation of cytR. We show that cytR expression is negatively controlled by the CytR protein and positively affected by the cAMP/CAP complex. Footprinting studies with purified CAP protein revealed two CAP binding sites upstream of the cytR promoter. A previously described mutation (cytR*) in the cloned cytR gene, which results in the phenotypic suppression of a CytR operator mutation in the tsx P2 promoter, was analysed. DNA sequence analysis of the cytR* mutation revealed a G-C to an A-T base pair transition at position -34 bp relative to the translational initiation site of cytR. This point mutation activates a cryptic promoter that is stronger than the wild-type cytR promoter and leads to overproduction of the CytR repressor.
大肠杆菌cytR编码的阻遏蛋白(CytR)控制着几个参与核苷和脱氧核苷摄取及代谢的基因的表达。通过确定cytR基因的转录起始位点鉴定出了cytR启动子。分离出一个染色体cytR-lacZ +操纵子融合体并用于研究cytR的调控。我们发现cytR的表达受到CytR蛋白的负调控,并受到cAMP/CAP复合物的正影响。用纯化的CAP蛋白进行的足迹研究揭示了cytR启动子上游的两个CAP结合位点。对克隆的cytR基因中一个先前描述的突变(cytR*)进行了分析,该突变导致tsx P2启动子中CytR操纵子突变的表型抑制。cytR*突变的DNA序列分析显示,相对于cytR翻译起始位点,在-34 bp处发生了从G-C到A-T的碱基对转换。这个点突变激活了一个比野生型cytR启动子更强的隐蔽启动子,并导致CytR阻遏蛋白的过量产生。