Mukhopadhyay S, Schellhorn H E
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
J Bacteriol. 1994 Apr;176(8):2300-7. doi: 10.1128/jb.176.8.2300-2307.1994.
Escherichia coli produces two independently regulated hydroperoxidases (catalases) that protect the cell from toxic concentrations of hydrogen peroxide. Hydroperoxidase I (HPI) is induced by hydrogen peroxide in an OxyR-dependent manner, while hydroperoxidase II (HPII) synthesis is regulated by an alternative sigma factor called RpoS (KatF). The activities of both hydroperoxidases increase as exponentially growing cells enter stationary phase. In this study, we examined the growth phase-dependent expression of HPI. Treatment of early-exponential-phase cells with spent culture supernatant resulted in induction of HPI synthesis. Extracellular levels of hydrogen peroxide, accumulating in the culture supernatant during late exponential phase, were found to be lower than the concentrations normally required to induce OxyR-dependent synthesis of HPI. This finding suggested that factors other than hydrogen peroxide may play a role in HPI expression. Weak acids such as acetate, which accumulate in culture supernatant and have been implicated in the regulation of HPII, caused a sixfold increase in HPI expression. Increases in HPI synthesis, mediated by weak acids and spent culture fluid supernatant, could be prevented by chloramphenicol, indicating that de novo protein synthesis is required for induction. Expression studies using a plasmid-borne lacZ transcriptional fusion to katG, the structural gene for HPI, indicated that growth phase-dependent regulation of HPI occurs primarily at the level of transcription and is dependent on RpoS. These results suggest that there may be a common regulatory mechanism of HPI and HPII expression in addition to previously described independent control mechanisms.
大肠杆菌产生两种独立调控的氢过氧化物酶(过氧化氢酶),可保护细胞免受有毒浓度过氧化氢的伤害。氢过氧化物酶I(HPI)由过氧化氢以氧依赖的方式诱导产生,而氢过氧化物酶II(HPII)的合成则由一种名为RpoS(KatF)的替代西格玛因子调控。随着指数生长期的细胞进入稳定期,两种氢过氧化物酶的活性均呈指数增加。在本研究中,我们检测了HPI在不同生长阶段的表达情况。用培养上清液处理指数生长期早期的细胞,可诱导HPI的合成。发现在指数生长期后期,培养上清液中积累的细胞外过氧化氢水平低于正常诱导氧依赖型HPI合成所需的浓度。这一发现表明,除过氧化氢外的其他因素可能在HPI表达中起作用。诸如乙酸盐等弱酸会在培养上清液中积累,并且与HPII的调控有关,它可使HPI的表达增加六倍。由弱酸和培养上清液介导的HPI合成增加可被氯霉素阻止,这表明诱导需要从头合成蛋白质。使用与HPI的结构基因katG的质粒携带的lacZ转录融合进行的表达研究表明,HPI的生长阶段依赖性调控主要发生在转录水平,并且依赖于RpoS。这些结果表明,除了先前描述的独立控制机制外,HPI和HPII的表达可能存在共同的调控机制。