López-Sánchez Aroa, Rivas-Marín Elena, Martínez-Pérez Olga, Floriano Belén, Santero Eduardo
Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide/CSIC, Carretera de Utrera, Sevilla, Spain.
Mol Microbiol. 2009 Sep;73(6):1086-100. doi: 10.1111/j.1365-2958.2009.06834.x. Epub 2009 Aug 4.
The genes required for tetralin biodegradation by Sphingomonas macrogolitabida strain TFA are clustered in two divergent and closely linked operons. ThnR, a LysR-type regulator, activates transcription from each operon in response to tetralin. The regulatory thnR gene is co-transcribed with the catabolic genes thnC, thnA3 and thnA4, resulting in positive autoregulation. ThnR binds with different affinity to two primary binding sites, designated B and C, in the intervening region between the two operons and makes additional contact with secondary sites that extend towards the promoters. In addition, ThnR may interact with itself when bound to each site via the formation of a DNA loop, as evidenced by the distortion of the DNA between the primary binding sites and the elimination of the higher-order complexes following the introduction of a half-turn of the DNA helix between the primary binding sites. Transcription from each promoter is not fully independent since mutations in each binding site affected transcription from both promoters. Based on these results, we propose a model of transcription activation that involves the formation of a complex structure by interactions between ThnR molecules bound to distant binding sites and favours transcription from one promoter to the detriment of the other.
嗜麦芽寡养单胞菌TFA菌株对四氢萘进行生物降解所需的基因聚集在两个反向且紧密相连的操纵子中。LysR型调控因子ThnR响应四氢萘激活每个操纵子的转录。调控基因thnR与分解代谢基因thnC、thnA3和thnA4共同转录,从而实现正自调控。ThnR以不同亲和力与两个操纵子之间间隔区域的两个主要结合位点(称为B和C)结合,并与向启动子延伸的次要位点进行额外接触。此外,ThnR在与每个位点结合时可能通过形成DNA环而自身相互作用,这一点可由主要结合位点之间的DNA扭曲以及在主要结合位点之间引入半圈DNA螺旋后高阶复合物的消失得到证明。每个启动子的转录并非完全独立,因为每个结合位点的突变都会影响两个启动子的转录。基于这些结果,我们提出了一种转录激活模型,该模型涉及通过与远距离结合位点结合的ThnR分子之间的相互作用形成复杂结构,并且有利于一个启动子的转录而不利于另一个启动子的转录。