Willett Jonathan, Smart James L, Bauer Carl E
Department of Biology, Jordan Hall, Bloomington, IN 47405, USA.
J Bacteriol. 2007 Nov;189(21):7765-73. doi: 10.1128/JB.00853-07. Epub 2007 Jul 6.
We provide in vivo genetic and in vitro biochemical evidence that RegA directly regulates bacteriochlorophyll and carotenoid biosynthesis in Rhodobacter capsulatus. beta-Galactosidase expression assays with a RegA-disrupted strain containing reporter plasmids for Mg-protoporphyrin IX monomethyl ester oxidative cyclase (bchE), Mg-protoporphyrin IX chelatase (bchD), and phytoene dehydrogenase (crtI) demonstrate RegA is responsible for fourfold anaerobic induction of bchE, threefold induction of bchD, and twofold induction of crtI. Promoter mapping studies, coupled with DNase I protection assays, map the region of RegA binding to three sites in the bchE promoter region. Similar studies at the crtA and crtI promoters indicate that RegA binds to a single region equidistant from these divergent promoters. These results demonstrate that RegA is directly responsible for anaerobic induction of bacteriochlorophyll biosynthesis genes bchE, bchD, bchJ, bchI, bchG, and bchP and carotenoid biosynthesis genes crtI, crtB, and crtA.
我们提供了体内遗传学和体外生物化学证据,证明RegA直接调控荚膜红细菌中细菌叶绿素和类胡萝卜素的生物合成。使用含有镁原卟啉IX单甲酯氧化环化酶(bchE)、镁原卟啉IX螯合酶(bchD)和八氢番茄红素脱氢酶(crtI)报告质粒的RegA缺失菌株进行的β-半乳糖苷酶表达分析表明,RegA负责bchE的四倍厌氧诱导、bchD的三倍诱导和crtI的两倍诱导。启动子图谱研究与DNase I保护分析相结合,将RegA结合区域定位到bchE启动子区域的三个位点。在crtA和crtI启动子上的类似研究表明,RegA结合到与这些不同启动子等距的单个区域。这些结果表明,RegA直接负责厌氧诱导细菌叶绿素生物合成基因bchE、bchD、bchJ、bchI、bchG和bchP以及类胡萝卜素生物合成基因crtI、crtB和crtA。