Rivas-Marín Elena, Floriano Belén, Santero Eduardo
Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide/Consejo Superior de Investigaciones Científicas/Junta de Andalucía, and Departamento de Biología Molecular e Ingeniería Bioquímica, Universidad Pablo de Olavide, Spain.
Sci Rep. 2016 Apr 18;6:24538. doi: 10.1038/srep24538.
Regulation of tetralin biodegradation operons is one of the examples of unconventional LysR-type mediated transcriptional regulation. ThnR activates transcription from two divergent and closely located promoters PB and PC. Although ThnR activates each promoter independently, transcription from each one increases when both promoters are together. Mutational analysis of the intergenic region shows that cooperative transcription is achieved through formation of a ThnR complex when bound to its respective sites at each promoter, via formation of a DNA loop. Mutations also defined ThnR contact sites that are important for independent transcriptional activation at each promoter. A mutation at the PB promoter region, which abolishes its independent transcription, does not affect at all PB transcription in the presence of the divergent promoter PC, thus indicating that the complex formed via DNA loop can compensate for the deficiencies in the correct protein-DNA interaction at one of the promoters. Combination of mutations in both promoters identifies a region at PC that is not important for its independent transcription but it is essential for cooperative transcription from both promoters. This work provides new insights into the diversity and complexity of activation mechanisms used by the most abundant type of bacterial transcriptional regulators.
四氢萘生物降解操纵子的调控是非常规赖斯R型介导的转录调控的例子之一。ThnR激活来自两个方向相反且位置紧密的启动子PB和PC的转录。尽管ThnR独立激活每个启动子,但当两个启动子同时存在时,每个启动子的转录都会增加。对基因间区域的突变分析表明,协同转录是通过ThnR复合物在每个启动子与其各自位点结合时形成DNA环来实现的。突变还确定了对每个启动子独立转录激活很重要的ThnR接触位点。PB启动子区域的一个突变消除了其独立转录,但在存在方向相反的启动子PC时,对PB转录完全没有影响,这表明通过DNA环形成的复合物可以弥补其中一个启动子上正确的蛋白质 - DNA相互作用的缺陷。两个启动子中的突变组合确定了PC上的一个区域,该区域对其独立转录不重要,但对两个启动子的协同转录至关重要。这项工作为最丰富类型的细菌转录调节因子所使用的激活机制的多样性和复杂性提供了新的见解。