Ivanova A B, Glinsky G V, Eisenstark A
Cancer Research Center, University of Missouri, Columbia 65201, USA.
Free Radic Biol Med. 1997;23(4):627-36. doi: 10.1016/s0891-5849(97)00013-0.
Escherichia coli delta oxyR mutants are hyper-sensitive to oxidative agents but this sensitivity is reversed to hyper-resistance in delta oxyR suppressor strains (delta oxyRsup; Greenberg, J.T. and Demple, B. 1988. EMBO J. 7:2611-2618). Also, delta oxyR mutants have increased mutation rates that are also reversed in delta oxyRsup. We now report that the rpoS regulon may have a role in determining hyper-resistance and loss of hyper-mutability of delta oxyRsup. Delta oxyRsup cells were also resistant to near-ultraviolet radiation (near-UV) and survived longer in stationary phase than delta oxyR cells. In delta oxyRsup cells elevated beta-galactosidase expression from a rpoS::lacZ promoter fusion and significant overproduction of RpoS protein was observed. These increases were accompanied by substantial elevation in transcription of rpoS-dependent genes as determined by beta-galactosidase expression from katE::lacZ, dps::lacZ, and xthA::lacZ promoters. Catalase HPI and HPII activities were also increased. When rpoS::Tn10 was transduced into delta oxyRsup, phenotypes switched back to hyper-sensitive, hyper-mutable and reduced catalases I and II. Individual delta oxyR colonies exhibited significant clonal variability in beta-galactosidase expression from rpoS::lacZ promoter. These results provide further evidence of the functional and regulatory overlap between two major anti-oxidant defense systems of bacteria.
大肠杆菌oxyR缺失突变体对氧化试剂高度敏感,但在oxyR缺失抑制菌株(oxyRsup;格林伯格,J.T.和登普尔,B. 1988. 《欧洲分子生物学组织杂志》7:2611 - 2618)中这种敏感性会转变为高度抗性。此外,oxyR缺失突变体的突变率增加,在oxyRsup中也会逆转。我们现在报告,rpoS调控子可能在决定oxyRsup的高度抗性和高突变性丧失方面发挥作用。oxyRsup细胞对近紫外辐射(近紫外)也具有抗性,并且在稳定期比oxyR细胞存活时间更长。在oxyRsup细胞中,观察到来自rpoS::lacZ启动子融合的β - 半乳糖苷酶表达升高以及RpoS蛋白大量过量产生。这些增加伴随着rpoS依赖性基因转录的显著升高,这是通过来自katE::lacZ、dps::lacZ和xthA::lacZ启动子的β - 半乳糖苷酶表达来确定的。过氧化氢酶HPI和HPII的活性也增加。当rpoS::Tn10转导到oxyRsup中时,表型又变回高度敏感、高突变性以及过氧化氢酶I和II减少。单个oxyR菌落从rpoS::lacZ启动子的β - 半乳糖苷酶表达表现出显著的克隆变异性。这些结果为细菌的两个主要抗氧化防御系统之间的功能和调控重叠提供了进一步的证据。