Dong Chen, Elsen Sylvie, Swem Lee R, Bauer Carl E
Department of Biology, Jordan Hall, Indiana University, Bloomington, IN 47405, USA.
J Bacteriol. 2002 May;184(10):2805-14. doi: 10.1128/JB.184.10.2805-2814.2002.
Open reading frame orf192, which is located immediately upstream of the aerobic repressor gene crtJ, was genetically and biochemically demonstrated to code for a second aerobic repressor (AerR) of photosynthesis gene expression in Rhodobacter capsulatus. Promoter-mapping studies indicate that crtJ has its own promoter but that a significant proportion of crtJ expression is promoted by read-through transcription of orf192 (aerR) transcripts through crtJ. Disruption of aerR resulted in increased photopigment biosynthesis during aerobic growth to a level similar to that of disruption of crtJ. Like that reported for CrtJ, beta-galactosidase assays of reporter gene expression indicated that disruption of aerR resulted in a two- to threefold increase in aerobic expression of the crtI and pucB operons. However, unlike CrtJ, AerR aerobically represses puf operon expression and does not aerobically repress bchC expression. Gel mobility shift analysis with purified AerR indicates that AerR does not bind to a bchC promoter probe but does bind to the crtI, puc, and puf promoter probes. These results indicate that AerR is a DNA-binding protein that targets genes partially overlapping a subset of genes that are also controlled by CrtJ. We also provide evidence for cooperative binding of AerR and CrtJ to the puc promoter region.
开放阅读框orf192位于需氧阻遏蛋白基因crtJ的紧邻上游位置,通过遗传学和生物化学方法证明其编码荚膜红细菌光合作用基因表达的第二种需氧阻遏蛋白(AerR)。启动子图谱研究表明,crtJ有其自身的启动子,但crtJ的很大一部分表达是由orf192(aerR)转录本通过crtJ的通读转录所促进的。aerR的破坏导致需氧生长期间光色素生物合成增加,达到与crtJ破坏相似的水平。与报道的CrtJ情况一样,报告基因表达的β-半乳糖苷酶分析表明,aerR的破坏导致crtI和pucB操纵子的需氧表达增加两到三倍。然而,与CrtJ不同的是,AerR在需氧条件下抑制puf操纵子的表达,而在需氧条件下不抑制bchC的表达。用纯化的AerR进行凝胶迁移率变动分析表明,AerR不与bchC启动子探针结合,但与crtI、puc和puf启动子探针结合。这些结果表明,AerR是一种DNA结合蛋白,其靶向的基因与也受CrtJ控制的一部分基因部分重叠。我们还提供了AerR和CrtJ协同结合到puc启动子区域的证据。