Parsot C, Mekalanos J J
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115.
Proc Natl Acad Sci U S A. 1990 Dec;87(24):9898-902. doi: 10.1073/pnas.87.24.9898.
The toxR gene of Vibrio cholerae encodes a transmembrane, DNA-binding protein that positively controls transcription of the genes for cholera toxin, TCP pili, and other proteins important in cholera pathogenesis. Nucleotide sequence analysis of the toxR upstream region has revealed that the heat shock gene htpG, encoding the bacterial homologue of the eukaryotic Hsp90 protein, was located immediately upstream and was divergently transcribed from toxR. Using lacZ transcriptional fusions, we have shown that neither toxR nor htpG expression was regulated by ToxR. However, the growth temperature had a coordinate but reciprocal effect on the expression from both the toxR and htpG promoters in V. cholerae; the decrease of toxR expression between 22 degrees C and 37 degrees C was proportional to the increase of htpG expression observed within that temperature range. A similar pattern of expression of the htpG and toxR promoters was observed in the heterologous host Escherichia coli, where this regulation was controlled by the level of the E. coli rpoH (htpR) gene product, sigma-32. Consistent with the temperature-regulated expression of the V. cholerae htpG promoter in E. coli, a sequence similar to the consensus sequence of the E. coli heat shock promoters was detected upstream from the V. cholerae htpG gene. We propose a model in which the regulation of toxR expression by temperature is controlled by the level of sigma-32 (RpoH) RNA polymerase.
霍乱弧菌的toxR基因编码一种跨膜DNA结合蛋白,该蛋白正向调控霍乱毒素、TCP菌毛及其他在霍乱发病机制中起重要作用的蛋白质的基因转录。对toxR上游区域的核苷酸序列分析显示,编码真核Hsp90蛋白细菌同源物的热休克基因htpG紧邻toxR上游,且与toxR呈反向转录。利用lacZ转录融合技术,我们发现toxR和htpG的表达均不受ToxR调控。然而,生长温度对霍乱弧菌中toxR和htpG启动子的表达具有协同但相反的影响;在22℃至37℃之间,toxR表达的降低与该温度范围内观察到的htpG表达的增加成比例。在异源宿主大肠杆菌中也观察到了htpG和toxR启动子的类似表达模式,其中这种调控由大肠杆菌rpoH(htpR)基因产物σ-32的水平控制。与大肠杆菌中霍乱弧菌htpG启动子的温度调控表达一致,在霍乱弧菌htpG基因上游检测到一个与大肠杆菌热休克启动子共有序列相似的序列。我们提出了一个模型,其中温度对toxR表达的调控由σ-32(RpoH)RNA聚合酶的水平控制。