Susa Motoki, Kubori Tomoko, Shimamoto Nobuo
Structural Biology Center, National Institute of Genetics, The Graduate University for Advanced Studies, Mishima, Shizuoka, Japan.
Mol Microbiol. 2006 Mar;59(6):1807-17. doi: 10.1111/j.1365-2958.2006.05058.x.
In transcription initiation, all RNA polymerase molecules bound to a promoter have been conventionally supposed to proceed into elongation of transcript. However, for Escherichia coli RNA polymerase, evidence has been accumulated for a view that only its fraction can proceed into elongation and the rest is retained at a promoter in non-productive form: a pathway branching in transcription initiation. Proteins such as GreA and GreB affect these fractions at several promoters in vitro. To reveal the ubiquitous existence of the branched mechanism in E. coli, we searched for candidate genes whose transcription decreased by disruption of greA and greB using a DNA array. Among the arbitrarily selected 11 genes from over 100, the atpC, cspA and rpsA passed the test by Northern blotting. The Gre factors activated transcription initiation from their promoters in vitro, and the results demonstrated that the branched mechanism is exploited in vivo regulation. Consistently, decrease in the level of the GreA in an anaerobic stationary condition accompanied a decrease in the levels of transcripts of these genes.
在转录起始过程中,传统上认为所有与启动子结合的RNA聚合酶分子都会进入转录延伸阶段。然而,对于大肠杆菌RNA聚合酶,有证据支持这样一种观点,即只有一部分能够进入延伸阶段,其余部分以非生产性形式保留在启动子处:这是一种在转录起始阶段分支的途径。诸如GreA和GreB等蛋白质在体外会影响多个启动子处的这些比例。为了揭示大肠杆菌中这种分支机制的普遍存在,我们使用DNA阵列寻找那些因greA和greB缺失而转录减少的候选基因。在从100多个基因中随机选择的11个基因中,atpC、cspA和rpsA通过Northern印迹法测试。Gre因子在体外激活了它们启动子的转录起始,结果表明这种分支机制被用于体内调控。同样,在厌氧静止条件下GreA水平的降低伴随着这些基因转录本水平的降低。