Krieger R, Rompf A, Schobert M, Jahn D
Institut für Organische Chemie und Biochemie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg i. Br., Germany.
Mol Genet Genomics. 2002 May;267(3):409-17. doi: 10.1007/s00438-002-0672-7. Epub 2002 Apr 26.
Pseudomonas aeruginosa forms most of its heme under anaerobic denitrifying conditions. To study the regulation of the hemA gene, which codes for the first enzyme of heme biosynthesis in P. aeruginosa, a lacZ reporter gene fusion was constructed. Expression of lacZ under the control of the hemA promoter was found to be increased by 2.8-fold under anaerobic conditions in the presence of the alternative electron acceptor nitrate, relative to the level observed under aerobic growth conditions. Anaerobic fermentative growth or the presence of nitrite did not affect the lacZ expression. The genes encoding the oxygen sensor protein Anr, the redox regulator Dnr, the nitrate regulator NarL and the DNA-bending Integration Host Factor (IHF) are all required for the cooperative anaerobic induction of the hemA promoter hemAp (1). Potential binding sites for these regulatory proteins were identified by site-directed mutagenesis of the promoter fused to the reporter gene. The mode of regulation of P. aeruginosa hemA differs significantly from that described for the hemA gene of Escherichia coli K-12.
铜绿假单胞菌在厌氧反硝化条件下合成大部分血红素。为了研究编码铜绿假单胞菌血红素生物合成首个酶的hemA基因的调控机制,构建了一个lacZ报告基因融合体。发现在厌氧条件下,当存在替代电子受体硝酸盐时,在hemA启动子控制下的lacZ表达相对于有氧生长条件下观察到的水平增加了2.8倍。厌氧发酵生长或亚硝酸盐的存在不影响lacZ表达。编码氧传感器蛋白Anr、氧化还原调节因子Dnr、硝酸盐调节因子NarL和DNA弯曲整合宿主因子(IHF)的基因都是hemA启动子hemAp厌氧协同诱导所必需的(1)。通过对与报告基因融合的启动子进行定点诱变,确定了这些调节蛋白的潜在结合位点。铜绿假单胞菌hemA的调控模式与大肠杆菌K-12的hemA基因所描述的调控模式有显著差异。