Bohannon D E, Connell N, Keener J, Tormo A, Espinosa-Urgel M, Zambrano M M, Kolter R
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
J Bacteriol. 1991 Jul;173(14):4482-92. doi: 10.1128/jb.173.14.4482-4492.1991.
Many of the changes in gene expression observed when Escherichia coli cells enter stationary phase are regulated at the level of transcription initiation. A group of stationary-phase-inducible promoters, known as "gearbox" promoter, display a characteristic sequence in the -10 region which differs greatly from the consensus sequence for sigma 70-dependent promoters. Here we describe our studies on the gearbox promoters bolAp1 and mcbAp, responsible for the temporally regulated transcription of bolA and the genes involved in the synthesis of the peptide antibiotic microcin B17, respectively. Deletion analysis of mcbAp demonstrated that the stationary-phase-inducible properties of this promoter are found in a DNA fragment extending from -54 to +11 bp, surrounding the transcriptional start site, and are separable from DNA sequences responsible for the OmpR-dependent stimulation of transcription of mcbAp. In vitro transcription studies indicate that the RNA polymerase holoenzyme involved in the transcription of mcbAp contains sigma 70. In this and an accompanying paper (R. Lange and R. Hengge-Aronis, J. Bacteriol. 173: 4474-4481, 1991), experiments are described which show that the product of katF, a global regulator of stationary-phase gene expression and a putative sigma factor, is required for the expression of bolAp1 fused to the reporter gene lacZ. In contrast, mcbAp appears to be negatively regulated by katF. We discuss the implications of these results for postexponential gene expression and the role of gearbox sequences in the regulation of promoter activity.
当大肠杆菌细胞进入稳定期时,所观察到的许多基因表达变化是在转录起始水平受到调控的。一组被称为“变速箱”启动子的稳定期诱导型启动子,在其-10区域显示出一种特征性序列,该序列与依赖于σ70的启动子的共有序列有很大差异。在此,我们描述了对变速箱启动子bolAp1和mcbAp的研究,它们分别负责bolA的时间调控转录以及与肽抗生素微菌素B17合成相关的基因的转录。对mcbAp的缺失分析表明,该启动子的稳定期诱导特性存在于一个从-54到+11 bp的DNA片段中,该片段围绕转录起始位点,并且与负责OmpR依赖性刺激mcbAp转录的DNA序列是可分离的。体外转录研究表明,参与mcbAp转录的RNA聚合酶全酶含有σ70。在本文以及一篇配套论文(R. Lange和R. Hengge-Aronis,《细菌学杂志》173: 4474 - 4481,1991)中,描述了一些实验,这些实验表明katF的产物是一种稳定期基因表达的全局调节因子和一种假定的σ因子,它是与报告基因lacZ融合的bolAp1表达所必需的。相反,mcbAp似乎受到katF的负调控。我们讨论了这些结果对指数后期基因表达的影响以及变速箱序列在启动子活性调控中的作用。