Conlin C A, Trun N J, Silhavy T J, Miller C G
Department of Microbiology, University of Illinois, Urbana-Champaign 61801.
J Bacteriol. 1992 Sep;174(18):5881-7. doi: 10.1128/jb.174.18.5881-5887.1992.
Mutations at the Escherichia coli prlC locus suppress the export defect of certain lamB signal sequence mutations. The Salmonella typhimurium opdA gene encodes an endoprotease that can participate in the catabolism of certain peptides and is required for normal development of phage P22. Plasmids carrying either the wild-type (pTC100 prlC+) or suppressor alleles of prlC complemented all phenotypes associated with an S. typhimurium opdA mutation. A plasmid carrying an amber mutation in prlC [prlC31(AM)] was unable to complement except in an amber suppressor background. Tn1000 insertions which eliminated the ability of pTC100 (prlC+) to complement opdA mapped to the region of the plasmid shown by deletion analysis and subcloning to contain prlC. The nucleotide sequence of a 2.7-kb fragment including this region was determined, revealing an open reading frame encoding a 77-kDa protein. The sequences of this open reading frame and its putative promoter region were very similar (84% nucleotide sequence identity and 95% amino acid identity) to those of S. typhimurium opdA, showing that these genes are homologs. The nucleotide sequence of the prlC1 suppressor allele was determined and predicts an in-frame duplication of seven amino acids, providing further confirmation that the prlC suppressor phenotype results from changes in the endopeptidase OpdA.
大肠杆菌prlC位点的突变可抑制某些lamB信号序列突变的输出缺陷。鼠伤寒沙门氏菌opdA基因编码一种内切蛋白酶,该酶可参与某些肽的分解代谢,并且是噬菌体P22正常发育所必需的。携带prlC野生型(pTC100 prlC+)或抑制等位基因的质粒可互补与鼠伤寒沙门氏菌opdA突变相关的所有表型。携带prlC琥珀突变[prlC31(AM)]的质粒无法互补,除非在琥珀抑制背景下。Tn1000插入消除了pTC100(prlC+)互补opdA的能力,其插入位点映射到通过缺失分析和亚克隆显示包含prlC的质粒区域。确定了包含该区域的2.7kb片段的核苷酸序列,揭示了一个编码77kDa蛋白质的开放阅读框。该开放阅读框及其推定启动子区域的序列与鼠伤寒沙门氏菌opdA的序列非常相似(核苷酸序列同一性为84%,氨基酸同一性为95%),表明这些基因是同源物。确定了prlC1抑制等位基因的核苷酸序列,并预测了七个氨基酸的框内重复,进一步证实了prlC抑制表型是由内肽酶OpdA的变化引起的。