McEvoy J L, Murata H, Chatterjee A K
Department of Plant Pathology, University of Missouri, Columbia 65211.
J Bacteriol. 1992 Aug;174(16):5471-4. doi: 10.1128/jb.174.16.5471-5474.1992.
In Erwinia carotovora subsp. carotovora 71, the induction of pectin lyase (Pnl), the bacteriocin carotovoricin (Ctv), and cellular lysis (Lss) requires a RecA function. We obtained mutants wherein a pleiotropic defect, i.e., the lack of induction with mitomycin C, is not restored by the recA+ DNA. From a genomic library of strain 71, a cosmid (pAKC280) that restored induction of Pnl, Ctv, and Lss by mitomycin C was isolated. The activator function, designated Rdg for regulator of damage-inducible genes, was localized by subcloning and insertional mutagenesis to a 2.6-kb region within a 6.7-kb EcoRI fragment. An rdg-lacZ operon fusion was inducible by mitomycin C in RecA+ but not RecA- derivatives of E. carotovora subsp. carotovora 71 and Escherichia coli. A RecA+ E. coli strain carrying only a PnlA+ plasmid was not inducible for Pnl production; however, when both a PnlA+ plasmid and a Rdg+ plasmid were present, the transcription of pnlA and the production of the enzyme were activated by mitomycin C. The size of the pnlA transcript produced in E. coli was identical to that of the transcript produced by E. carotovora subsp. carotovora 71, suggesting that the same promoter and termination sequences were being utilized in these bacteria.
在胡萝卜软腐欧文氏菌胡萝卜软腐亚种71中,果胶裂解酶(Pnl)、细菌素胡萝卜软腐菌素(Ctv)的诱导以及细胞裂解(Lss)需要RecA功能。我们获得了一些突变体,其中多效性缺陷,即对丝裂霉素C缺乏诱导作用,不能被recA⁺ DNA恢复。从菌株71的基因组文库中,分离出一个通过丝裂霉素C恢复Pnl、Ctv和Lss诱导作用的黏粒(pAKC280)。通过亚克隆和插入诱变将指定为Rdg(损伤诱导基因调节因子)的激活功能定位到一个6.7 kb EcoRI片段内的2.6 kb区域。rdg-lacZ操纵子融合在胡萝卜软腐欧文氏菌胡萝卜软腐亚种71和大肠杆菌的RecA⁺衍生物中可被丝裂霉素C诱导,但在RecA⁻衍生物中不能被诱导。仅携带PnlA⁺质粒的RecA⁺大肠杆菌菌株不能被诱导产生Pnl;然而,当同时存在PnlA⁺质粒和Rdg⁺质粒时,pnlA的转录和该酶的产生可被丝裂霉素C激活。在大肠杆菌中产生的pnlA转录本的大小与胡萝卜软腐欧文氏菌胡萝卜软腐亚种71产生的转录本大小相同,这表明这些细菌利用了相同的启动子和终止序列。