Davies Karen M, Dedman Amy J, van Horck Stephanie, Lewis Peter J
School of Environmental and Life Sciences, Biological Sciences, University of Newcastle, Callaghan, NSW 2308, Australia.
Mol Microbiol. 2005 Jul;57(2):366-79. doi: 10.1111/j.1365-2958.2005.04469.x.
Bacterial RNA polymerases (RNAPs) are capable of producing full-length transcripts in the absence of additional factors using in vitro assays. However, in vivo RNAP can become stalled during the elongation phase of transcription due to the presence of various sequence motifs. Subsequently, a host of elongation factors are required to modulate the activity of RNAP. NusA, the most intensively studied elongation factor, plays a role in increasing RNAP pausing and termination. Conversely, it is also important in transcription of rRNA where it functions as an anti-termination factor, helping to ensure only full-length transcripts are produced. Here we show that NusA is closely associated with RNAP within the bacterial nucleoid and that it is preferentially recruited to sites of rRNA synthesis. In vivo and in vitro analyses indicate this results in a change in stoichiometry of NusA:RNAP from 1:1 to approximately 2:1 at the subcellular sites of rRNA synthesis. A model is presented showing how the ratio of NusA:RNAP could affect the activity of the elongation complex so that it functions as an anti-terminator complex during rRNA synthesis.
细菌RNA聚合酶(RNAPs)能够在体外测定中在没有其他因子的情况下产生全长转录本。然而,在体内,由于各种序列基序的存在,RNAP在转录延伸阶段可能会停滞。随后,需要一系列延伸因子来调节RNAP的活性。NusA是研究最深入的延伸因子,在增加RNAP暂停和终止方面发挥作用。相反,它在rRNA转录中也很重要,在那里它作为抗终止因子发挥作用,有助于确保只产生全长转录本。在这里,我们表明NusA与细菌类核中的RNAP紧密相关,并且它优先被招募到rRNA合成位点。体内和体外分析表明,这导致在rRNA合成的亚细胞位点上NusA:RNAP的化学计量比从1:1变为约2:1。本文提出了一个模型,展示了NusA:RNAP的比例如何影响延伸复合物的活性,使其在rRNA合成过程中作为抗终止复合物发挥作用。