Liu Y, Chatterjee A, Chatterjee A K
Department of Plant Pathology, University of Missouri, Columbia 65211.
Mol Microbiol. 1994 Dec;14(5):999-1010. doi: 10.1111/j.1365-2958.1994.tb01334.x.
In most soft-rotting Erwinia spp., including E. carotovora subsp. carotovora strain 71 (Ecc71), production of the plant cell wall degrading enzyme pectin lyase (Pnl) is activated by DNA-damaging agents such as mitomycin C (MC). Induction of Pnl production in Ecc71 requires a functional recA gene and the rdg locus. DNA sequencing and RNA analyses revealed that the rdg locus contains two regulatory genes, rdgA and rdgB, in separate transcriptional units. There is high homology between RdgA and repressors of lambdoid phages, specially phi 80. RdgB, however, has significant homology with transcriptional activators of Mu phage. Both RdgA and RdgB are also predicted to possess helix-turn-helix motifs. By replacing the rdgB promoter with the IPTG-inducible tac promoter, we have determined that rdgB by itself can activate Pnl production in Escherichia coli. However, deletion analysis of rdg+ DNA indicated that, when driven by their native promoters, functions of both rdgA and rdgB are required for the induction of pnlA expression by MC treatment. While rdgB transcription occurs only after MC treatment, a substantial level of rdgA mRNA is detected in the absence of MC treatment. Moreover, upon induction with MC, a new rdgA mRNA species, initiated from a different start site, is produced at a high level. Thus, the two closely linked rdgA and rdgB genes, required for the regulation of Pnl production, are expressed differently in Ecc71.
在大多数软腐欧文氏菌属细菌中,包括胡萝卜软腐欧文氏菌胡萝卜软腐亚种71菌株(Ecc71),植物细胞壁降解酶果胶裂解酶(Pnl)的产生会被诸如丝裂霉素C(MC)等DNA损伤剂激活。在Ecc71中诱导Pnl产生需要一个功能性的recA基因和rdg位点。DNA测序和RNA分析表明,rdg位点在不同的转录单元中包含两个调控基因,rdgA和rdgB。RdgA与λ样噬菌体的阻遏物,特别是φ80,具有高度同源性。然而,RdgB与Mu噬菌体的转录激活因子具有显著同源性。RdgA和RdgB也都被预测具有螺旋-转角-螺旋基序。通过用IPTG诱导型tac启动子替换rdgB启动子,我们已经确定rdgB自身就能在大肠杆菌中激活Pnl的产生。然而,对rdg+ DNA的缺失分析表明,当由其天然启动子驱动时,MC处理诱导pnlA表达需要rdgA和rdgB的功能。虽然rdgB转录仅在MC处理后发生,但在未进行MC处理时能检测到相当水平的rdgA mRNA。此外,在用MC诱导后,从不同起始位点起始的一种新的rdgA mRNA种类会大量产生。因此,调控Pnl产生所需的两个紧密相连的rdgA和rdgB基因在Ecc71中的表达是不同的。