Schellhorn H E, Stones V L
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
J Bacteriol. 1992 Jul;174(14):4769-76. doi: 10.1128/jb.174.14.4769-4776.1992.
Chromosomal transcriptional and translational lacZ fusions to the katE (structural gene for the HPII hydroperoxidase) and katF (putative sigma factor required for katE expression) genes of Escherichia coli were isolated, and the regulation of these fusions was used to identify factors that control the expression of these two important antioxidant factors. While katE was found to be regulated primarily at the level of transcription (since induction patterns were similar for both transcriptional and translational fusions), katF expression was a function of both transcriptional and translational signals. The katE gene was induced 57-fold as cells entered the stationary phase, while katF was induced 23-fold. katF induction was coincident with katE induction and occurred at the onset of the stationary growth phase. Expression of both katE and katF could be induced by resuspending uninduced exponential-phase cells in spent culture supernatant recovered from stationary-phase cells. The component of stationary-phase culture supernatant responsible for induction of the katF regulon appeared to be acetate, since expression of both katE and katF fusions was induced when exponential-phase cells were exposed to this weak acid. Other weak acids, including propionate and benzoate, were also found to be effective inducers of expression of both katF and katE. Induction of katE and katF fusions was unaffected in merodiploid strains containing both mutant and wild-type alleles, indicating that expression of both genes is independent of the wild-type gene product. Examination of catalase zymograms prepared from cells exposed to various levels of acetate revealed that both HPI and HPII catalases are induced by this weak acid, suggesting that there is a common link in the regulation of these two enzymes.
分离出了大肠杆菌katE(HPII氢过氧化物酶的结构基因)和katF(katE表达所需的假定σ因子)基因的染色体转录和翻译lacZ融合体,并利用这些融合体的调控来鉴定控制这两种重要抗氧化因子表达的因素。虽然发现katE主要在转录水平受到调控(因为转录和翻译融合体的诱导模式相似),但katF的表达是转录和翻译信号共同作用的结果。随着细胞进入稳定期,katE基因被诱导了57倍,而katF被诱导了23倍。katF的诱导与katE的诱导同时发生,且发生在稳定生长期开始时。将未诱导的指数生长期细胞重悬于从稳定期细胞回收的用过的培养上清液中,可诱导katE和katF的表达。负责诱导katF调节子的稳定期培养上清液成分似乎是乙酸盐,因为当指数生长期细胞暴露于这种弱酸时,katE和katF融合体的表达都会被诱导。还发现其他弱酸,包括丙酸盐和苯甲酸盐,也是katF和katE表达的有效诱导剂。在同时含有突变和野生型等位基因的部分二倍体菌株中,katE和katF融合体的诱导不受影响,这表明这两个基因的表达不依赖于野生型基因产物。对暴露于不同水平乙酸盐的细胞制备的过氧化氢酶酶谱进行检查发现,HPI和HPII过氧化氢酶都被这种弱酸诱导,这表明这两种酶的调控存在共同联系。