Hirsch Matthew, Elliott Thomas
Department of Microbiology, Immunology, and Cell Biology, West Virginia University Health Science Center, Morgantown, WV 26506, USA.
J Bacteriol. 2005 Nov;187(21):7204-13. doi: 10.1128/JB.187.21.7204-7213.2005.
In enteric bacteria, adaptation to a number of different stresses is mediated by the RpoS protein, one of several sigma factors that collectively allow a tailored transcriptional response to environmental cues. Stress stimuli including low temperature, osmotic shock, nutrient limitation, and growth to stationary phase (SP) all result in a substantial increase in RpoS abundance and activity. The mechanism of regulation depends on the specific signal but may occur at the level of transcription, translation, protein activity, or targeted proteolysis. In both Escherichia coli and Salmonella enterica, SP induction of RpoS in rich medium is >30 fold and includes effects on both transcription and translation. Recently, we found that SP control of rpoS transcription in S. enterica involves repression of the major rpoS promoter during exponential phase by the global transcription factor Fis. Working primarily with E. coli, we now show that 24 nucleotides of the rpoS ribosome-binding site (RBS) are necessary and sufficient for a large part of the increase in rpoS translation as cells grow to SP. Genetic evidence points to an essential role for the leader nucleotides just upstream of the Shine-Dalgarno sequence but is conflicted on the question of whether sequence or structure is important. SP regulation of rpoS is conserved between E. coli and S. enterica. When combined with an fis mutation to block transcriptional effects, replacement of the rpoS RBS sequence by the lacZ RBS eliminates nearly all SP induction of RpoS.
在肠道细菌中,对多种不同应激的适应是由RpoS蛋白介导的,RpoS蛋白是几种σ因子之一,这些σ因子共同作用使得细菌能够根据环境线索进行定制化的转录反应。包括低温、渗透压休克、营养限制以及生长至稳定期(SP)在内的应激刺激,都会导致RpoS丰度和活性大幅增加。调控机制取决于特定信号,但可能发生在转录、翻译、蛋白质活性或靶向蛋白水解水平。在大肠杆菌和肠炎沙门氏菌中,在丰富培养基中SP诱导RpoS的倍数大于30倍,且对转录和翻译均有影响。最近,我们发现肠炎沙门氏菌中rpoS转录的SP调控涉及全局转录因子Fis在指数生长期对主要rpoS启动子的抑制。现在,我们主要以大肠杆菌为研究对象,发现随着细胞生长至稳定期,rpoS核糖体结合位点(RBS)的24个核苷酸对于rpoS翻译的大部分增加是必要且充分的。遗传学证据表明,Shine-Dalgarno序列上游的前导核苷酸起着至关重要的作用,但在序列或结构是否重要这一问题上存在争议。rpoS的SP调控在大肠杆菌和肠炎沙门氏菌之间是保守的。当与fis突变结合以阻断转录效应时,用lacZ RBS替换rpoS RBS序列几乎消除了RpoS的所有SP诱导。