Weston L A, Kadner R J
Department of Microbiology, School of Medicine, University of Virginia, Charlottesville 22908.
J Bacteriol. 1988 Aug;170(8):3375-83. doi: 10.1128/jb.170.8.3375-3383.1988.
The uhpABCT locus of Escherichia coli is responsible for expression of the sugar-phosphate transport system and its induction by external glucose 6-phosphate. Expression of uhpT-lacZ fusions depended on the function of uhpA, uhpB, and uhpC but not of uhpT. A plasmid carrying only uhpT conferred transport activity in a host strain deleted for the uhp region. Thus, uhpT encodes the polypeptide required for transport function, and the other three uhp genes regulate uhpT transcription. The presence of uhpA at elevated copy number resulted in a substantial increase in uhpT expression. This elevated expression was only about 50% of the level seen in induced haploid cells, and no further increase occurred after addition of inducer. Activation by multicopy uhpA was not affected by the status of uhpC but was decreased in the absence of uhpB, suggesting a role for UhpB in directly activating UhpA. Transcription of uhpA, monitored by expression of a uhpA-lacZ fusion, was not affected by either inducer or the presence of the wild-type uhpA allele. The presence of multiple copies of the uhpT promoter region reduced uhpT expression in strains with uhpA in single copy number but not in those with multiple copies, consistent with competition for the activator. Amino acid sequence comparisons showed that UhpA was homologous to a family of bacterial regulatory proteins, some of which act as transcriptional activators (OmpR, PhoB, NtrC, and DctD). The C-terminal portion of UhpB displayed matches to the corresponding portions of another family of proteins (EnvZ, PhoMR, NtrB, and DctB) that participate in regulation of gene expression in response to environmental factors.
大肠杆菌的uhpABCT基因座负责糖磷酸转运系统的表达及其由细胞外6-磷酸葡萄糖诱导的过程。uhpT - lacZ融合蛋白的表达依赖于uhpA、uhpB和uhpC的功能,而不依赖于uhpT的功能。仅携带uhpT的质粒在缺失uhp区域的宿主菌株中赋予转运活性。因此,uhpT编码转运功能所需的多肽,而其他三个uhp基因调节uhpT的转录。高拷贝数的uhpA导致uhpT表达大幅增加。这种升高的表达仅约为诱导单倍体细胞中观察到水平的50%,添加诱导剂后没有进一步增加。多拷贝uhpA的激活不受uhpC状态的影响,但在没有uhpB的情况下会降低,这表明UhpB在直接激活UhpA中起作用。通过uhpA - lacZ融合蛋白的表达监测,uhpA的转录不受诱导剂或野生型uhpA等位基因存在的影响。uhpT启动子区域的多个拷贝的存在降低了单拷贝uhpA菌株中的uhpT表达,但在多拷贝菌株中没有降低,这与对激活剂的竞争一致。氨基酸序列比较表明,UhpA与一类细菌调节蛋白同源,其中一些作为转录激活剂(OmpR、PhoB、NtrC和DctD)。UhpB的C末端部分与另一类蛋白质(EnvZ、PhoMR、NtrB和DctB)的相应部分匹配,这些蛋白质参与响应环境因素的基因表达调控。