Wong S L, Doi R H
J Biol Chem. 1984 Aug 10;259(15):9762-7.
The promoter region of Bacillus subtilis subtilisin E was found to be composed of two overlapping promoters with their transcription starting sites separated from each other by 15 base pairs (Wong, S.-L., Price, C. W., Goldfarb, D. S., and Doi, R. H. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 1184-1188). At least one of the promoters is transcribed by a minor form of B. subtilis RNA polymerase with a sigma factor of 37,000 daltons. In vitro transcription analyses and in vivo studies with promoter probe plasmids pKO-1 and pCED-6 demonstrated that Escherichia coli RNA polymerase was able to initiate transcription from the subtilisin promoter cluster. S1 nuclease-mapping studies with both in vivo and in vitro transcribed RNA from E. coli and B. subtilis illustrate that E. coli can initiate transcription from both promoters with the same transcription start points as B. subtilis. The promoter strength of this promoter cluster in E. coli, as expressed in terms of galactokinase units, was 64 units and represents weak promoter activity in the E. coli system. These data indicate that either the single E. coli RNA polymerase is able to recognize the minor sigma 37 promoter or E. coli contains a hitherto unrecognized minor RNA polymerase holoenzyme which is capable of recognizing a B. subtilis sigma 37 promoter. On the other hand the B. subtilis RNA polymerase holoenzymes have been quite promoter-specific in our experiments to date.
已发现枯草芽孢杆菌枯草杆菌蛋白酶E的启动子区域由两个重叠的启动子组成,它们的转录起始位点彼此相隔15个碱基对(Wong, S.-L., Price, C. W., Goldfarb, D. S., and Doi, R. H. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 1184 - 1188)。至少其中一个启动子由一种次要形式的枯草芽孢杆菌RNA聚合酶转录,该聚合酶带有一个37,000道尔顿的σ因子。用启动子探针质粒pKO - 1和pCED - 6进行的体外转录分析和体内研究表明,大肠杆菌RNA聚合酶能够从枯草杆菌蛋白酶启动子簇起始转录。对大肠杆菌和枯草芽孢杆菌体内和体外转录的RNA进行S1核酸酶图谱分析表明,大肠杆菌能够从两个启动子起始转录,转录起始点与枯草芽孢杆菌相同。以半乳糖激酶单位表示,该启动子簇在大肠杆菌中的启动子强度为64个单位,在大肠杆菌系统中代表较弱的启动子活性。这些数据表明,要么单一的大肠杆菌RNA聚合酶能够识别次要的σ37启动子,要么大肠杆菌含有一种迄今未被识别的次要RNA聚合酶全酶,它能够识别枯草芽孢杆菌的σ37启动子。另一方面,在我们目前的实验中,枯草芽孢杆菌RNA聚合酶全酶对启动子具有相当的特异性。