Shelver D, Kerby R L, He Y, Roberts G P
Department of Bacteriology, University of Wisconsin-Madison 53706.
J Bacteriol. 1995 Apr;177(8):2157-63. doi: 10.1128/jb.177.8.2157-2163.1995.
Induction of the CO-oxidizing system of the photosynthetic bacterium Rhodospirillum rubrum is regulated at the level of gene expression by the presence of CO. In this paper, we describe the identification of a gene that is required for CO-induced gene expression. An 11-kb deletion of the region adjacent to the previously characterized cooFSCTJ region resulted in a mutant unable to synthesize CO dehydrogenase in response to CO and unable to grow utilizing CO as an energy source. A 2.5-kb region that corresponded to a portion of the deleted region complemented this mutant for its CO regulation defect, restoring its ability to grow utilizing CO as an energy source. When the 2.5-kb region was sequenced, one open reading frame, designated cooA, predicted a product showing similarity to members of the cyclic AMP receptor protein (CRP) family of transcriptional regulators. The product, CooA, is 28% identical (51% similar) to CRP and 18% identical (45% similar) to FNR from Escherichia coli. The insertion of a drug resistance cassette into cooA resulted in a mutant that could not grow utilizing CO as an energy source. CooA contains a number of cysteine residues substituted at, or adjacent to, positions that correspond to residues that contact cyclic AMP in the crystal structure of CRP. A model based on this observation is proposed for the recognition of CO by Cooa. Adjacent to cooA are two genes, nadB and nadC, with predicted products similar to proteins in other bacteria that catalyze reactions in the de novo synthesis of NAD.(ABSTRACT TRUNCATED AT 250 WORDS)
光合细菌红螺菌(Rhodospirillum rubrum)的一氧化碳氧化系统的诱导在基因表达水平上受一氧化碳的存在调控。在本文中,我们描述了一个对一氧化碳诱导的基因表达所必需的基因的鉴定。与先前表征的cooFSCTJ区域相邻的区域发生11 kb的缺失,导致一个突变体,该突变体无法响应一氧化碳合成一氧化碳脱氢酶,也无法利用一氧化碳作为能源生长。一个与缺失区域的一部分相对应的2.5 kb区域弥补了该突变体的一氧化碳调节缺陷,恢复了其利用一氧化碳作为能源生长的能力。对该2.5 kb区域进行测序时,一个开放阅读框,命名为cooA,预测的产物与转录调节因子的环腺苷酸受体蛋白(CRP)家族成员具有相似性。该产物CooA与CRP有28%的同一性(51%的相似性),与大肠杆菌的FNR有18%的同一性(45%的相似性)。将一个抗药盒插入cooA导致一个突变体,该突变体无法利用一氧化碳作为能源生长。CooA含有一些半胱氨酸残基,这些残基在对应于CRP晶体结构中与环腺苷酸接触的残基的位置或其附近被取代。基于这一观察结果提出了一个CooA识别一氧化碳的模型。cooA相邻的是两个基因,nadB和nadC,其预测产物与其他细菌中催化NAD从头合成反应的蛋白质相似。(摘要截短于250字)