Raina S, Missiakas D, Baird L, Kumar S, Georgopoulos C
Department of Cellular, Viral and Molecular Biology, University of Utah School of Medicine, Salt Lake City 84132.
J Bacteriol. 1993 Aug;175(16):5009-21. doi: 10.1128/jb.175.16.5009-5021.1993.
We have characterized a new Escherichia coli operon consisting of two genes, ecpD and htrE. The ecpD gene encodes a 27-kDa protein which is 40% identical at the amino acid level to the pilin chaperone PapD family of proteins. Immediately downstream of the ecpD gene is the htrE gene. The htrE gene encodes a polypeptide of 95 kDa which is processed to a 92-kDa mature species. The HtrE protein is 38% identical to the type II pilin porin protein PapC. The ecpD htrE operon is located at 3.3 min on the genetic map, corresponding to the region from kbp 153 to 157 of the E. coli physical map. The htrE gene was identified on the basis of a Tn5 insertion mutation which resulted in a temperature-sensitive growth phenotype above 43.5 degrees C. The transcription of this operon is induced with a temperature shift from 22 to 37 or 42 degrees C but not to higher temperatures, e.g., 50 degrees C. Consistent with this result, the temperature-induced transcription was shown to be independent of the rpoH gene product (sigma 32). The transcription of this operon was further shown to require functional integration host factor protein, since himA or himD mutant bacteria possessed lower levels of ecpD htrE transcripts. Among the three transcriptional start sites discovered, one, defined by the P2 promoter, was found to be under the positive regulation of the katF (rpoS) gene, which encodes a putative sigma factor required for the transcription of many growth phase-regulated genes.
我们已鉴定出一个由ecpD和htrE两个基因组成的新型大肠杆菌操纵子。ecpD基因编码一种27 kDa的蛋白质,该蛋白质在氨基酸水平上与菌毛伴侣PapD家族蛋白质有40%的同一性。ecpD基因的紧邻下游是htrE基因。htrE基因编码一种95 kDa的多肽,该多肽经加工后成为92 kDa的成熟形式。HtrE蛋白与II型菌毛孔蛋白PapC有38%的同一性。ecpD htrE操纵子位于遗传图谱上的3.3分钟处,对应于大肠杆菌物理图谱上从153 kbp到157 kbp的区域。htrE基因是基于一个Tn5插入突变鉴定出来的,该突变导致在43.5℃以上出现温度敏感型生长表型。此操纵子的转录在温度从22℃转变至37℃或42℃时被诱导,但在更高温度(如50℃)下则不会。与该结果一致,温度诱导的转录显示与rpoH基因产物(σ32)无关。进一步研究表明,此操纵子的转录需要功能性整合宿主因子蛋白,因为himA或himD突变细菌中ecpD htrE转录本的水平较低。在发现的三个转录起始位点中,由P2启动子定义的一个位点被发现受katF(rpoS)基因的正调控,katF基因编码一种假定的σ因子,是许多生长阶段调控基因转录所必需的。