Sawers R Gary
Department of Molecular Microbiology, John Innes Centre, Norwich NR4 7UH, UK.
Mol Microbiol. 2005 Dec;58(5):1441-53. doi: 10.1111/j.1365-2958.2005.04915.x.
The anaerobically inducible dicistronic focA-pfl operon is transcribed from three co-ordinately regulated promoters that are located 5' of the operon. Remarkably, the 5' ends of four further highly abundant operon-internal transcripts are located within the focA gene, with a fifth transcript mapping in the intergenic region between focA and pfl. The findings of this study demonstrate that the bulk of these five operon-internal transcripts are the result of processing. Processing was independent of the broad-spectrum endoribonucleases associated with mRNA turnover and still occurred when the upstream regulatory region of the operon was replaced with two different heterologous promoters recognized by Escherichia coli core RNA polymerase, including the tetP promoter. However, when the T7Phi10 promoter was introduced upstream of the focA-pfl operon, mainly full-length transcript and a minor amount of two processing products were observed. T7 RNA polymerase mutants that exhibit reduced elongation speed did not restore the wild-type transcript-processing pattern. Moreover, processing was independent of focA translation. Taken together, these data suggest that processing of the focA-pfl transcripts occurs by a novel mechanism that might require the action of E. coli core RNA polymerase.
厌氧诱导型双顺反子focA - pfl操纵子由位于操纵子5'端的三个协同调控启动子转录。值得注意的是,另外四个高度丰富的操纵子内部转录本的5'端位于focA基因内,第五个转录本定位在focA和pfl之间的基因间区域。本研究结果表明,这五个操纵子内部转录本中的大部分是加工的结果。加工过程独立于与mRNA周转相关的广谱内切核糖核酸酶,并且当操纵子的上游调控区域被大肠杆菌核心RNA聚合酶识别的两种不同的异源启动子取代时(包括tetP启动子),加工过程仍然发生。然而,当在focA - pfl操纵子上游引入T7Phi10启动子时,主要观察到全长转录本和少量的两种加工产物。表现出延伸速度降低的T7 RNA聚合酶突变体并未恢复野生型转录本加工模式。此外,加工过程独立于focA翻译。综上所述,这些数据表明focA - pfl转录本的加工通过一种可能需要大肠杆菌核心RNA聚合酶作用的新机制发生。