Pogliano J, Lynch A S, Belin D, Lin E C, Beckwith J
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Genes Dev. 1997 May 1;11(9):1169-82. doi: 10.1101/gad.11.9.1169.
We show that the two-component signal transduction system of Escherichia coli, CpxA-CpxR, controls the expression of genes encoding cell envelope proteins involved in protein folding and degradation. These findings are based on three lines of evidence. First, activation of the Cpx pathway induces 5- to 10-fold the synthesis of DsbA, required for disulfide bond formation, and DegP, a major periplasmic protease. Second, using electrophoretic mobility shift and DNase I protection assays, we have shown that phosphorylated CpxR binds to elements upstream of the transcription start sites of dsbA, degP, and ppiA (rotA), the latter coding for a peptidyl-prolyl cis/trans isomerase. Third, we have demonstrated increased in vivo transcription of all three genes, dsbA, degP, and ppiA, when the Cpx pathway is activated. We have identified a putative CpxR consensus binding site that is found upstream of a number of other E. coli genes. These findings suggest a potentially extensive Cpx regulon including genes transcribed by sigma70 and sigma(E), which encode factors involved in protein folding as well as other cellular functions.
我们发现,大肠杆菌的双组分信号转导系统CpxA - CpxR可控制编码参与蛋白质折叠和降解的细胞包膜蛋白的基因的表达。这些发现基于三条证据。首先,Cpx途径的激活可使二硫键形成所需的DsbA以及主要的周质蛋白酶DegP的合成增加5至10倍。其次,通过电泳迁移率变动分析和DNase I保护分析,我们发现磷酸化的CpxR可与dsbA、degP和ppiA(rotA)转录起始位点上游的元件结合,后者编码一种肽基脯氨酰顺/反异构酶。第三,我们证明当Cpx途径被激活时,dsbA、degP和ppiA这三个基因在体内的转录均增加。我们鉴定出一个假定的CpxR共有结合位点,该位点存在于许多其他大肠杆菌基因的上游。这些发现表明存在一个潜在广泛的Cpx调控子,包括由sigma70和sigma(E)转录的基因,这些基因编码参与蛋白质折叠以及其他细胞功能的因子。