Arnvig Kristine B, Zeng Shirley, Quan Selwyn, Papageorge Alexander, Zhang Ning, Villapakkam Anuradha C, Squires Catherine L
Division of Mycobacterial Research, National Institute for Medical Research, London, UK.
J Bacteriol. 2008 Nov;190(21):7251-7. doi: 10.1128/JB.00760-08. Epub 2008 Aug 29.
Transcription antitermination in the ribosomal operons of Escherichia coli results in the modification of RNA polymerase by specific proteins, altering its basic properties. For such alterations to occur, signal sequences in rrn operons are required as well as individual interacting proteins. In this study we tested putative rrn transcription antitermination-inducing sequences from five different bacteria for their abilities to function in E. coli. We further examined their response to the lack of one known rrn transcription antitermination protein from E. coli, NusB. We monitored antitermination activity by assessing the ability of RNA polymerase to read through a factor-dependent terminator. We found that, in general, the closer the regulatory sequence matched that of E. coli, the more likely there was to be a successful antitermination-proficient modification of the transcription complex. The rrn leader sequences from Pseudomonas aeruginosa, Bacillus subtilis, and Caulobacter crescentus all provided various levels of, but functionally significant antitermination properties to, RNA polymerase, while those of Mycobacterium tuberculosis and Thermotoga maritima did not. Possible RNA folding structures of presumed antitermination sequences and specific critical bases are discussed in light of our results. An unexpected finding was that when using the Caulobacter crescentus rrn leader sequence, there was little effect on terminator readthrough in the absence of NusB. All other hybrid antitermination system activities required this factor. Possible reasons for this finding are discussed.
大肠杆菌核糖体操纵子中的转录抗终止作用会导致RNA聚合酶被特定蛋白质修饰,从而改变其基本特性。要发生这种改变,rrn操纵子中的信号序列以及单个相互作用蛋白是必需的。在本研究中,我们测试了来自五种不同细菌的假定rrn转录抗终止诱导序列在大肠杆菌中的功能。我们进一步研究了它们对大肠杆菌中一种已知的rrn转录抗终止蛋白NusB缺失的反应。我们通过评估RNA聚合酶通读因子依赖性终止子的能力来监测抗终止活性。我们发现,一般来说,调控序列与大肠杆菌的序列越匹配,转录复合物成功进行抗终止修饰的可能性就越大。铜绿假单胞菌、枯草芽孢杆菌和新月柄杆菌的rrn前导序列都为RNA聚合酶提供了不同程度但功能上显著的抗终止特性,而结核分枝杆菌和嗜热栖热菌的rrn前导序列则没有。根据我们的结果,讨论了假定抗终止序列可能的RNA折叠结构和特定关键碱基。一个意外的发现是,当使用新月柄杆菌的rrn前导序列时,在没有NusB的情况下对终止子通读几乎没有影响。所有其他杂交抗终止系统的活性都需要这个因子。讨论了这一发现的可能原因。