Fiorentino Gabriella, Ronca Raffaele, Cannio Raffaele, Rossi Mosè, Bartolucci Simonetta
Dipartimento di Biologia Strutturale e Funzionale, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S Angelo, Via Cinthia, 80126, Napoli, Italy.
J Bacteriol. 2007 Oct;189(20):7351-60. doi: 10.1128/JB.00885-07. Epub 2007 Aug 3.
A DNA binding protein, BldR, was identified in the crenarchaeon Sulfolobus solfataricus as a protein 5- to 10-fold more abundant in cells grown in the presence of toxic aldehydes; it binds to regulatory sequences located upstream of an alcohol dehydrogenase gene (Sso2536). BldR is homologous to bacterial representatives of the MarR (multiple antibiotic resistance) family of transcriptional regulators that mediate response to multiple environmental stresses. Transcriptional analysis revealed that the bldR gene was transcribed in a bicistronic unit composed of the genes encoding the transcriptional regulator (Sso1352) and a putative multidrug transporter (Sso1351) upstream. By homology to bacterial counterparts, the bicistron was named the mar-like operon. The level of mar-like operon expression was found to be increased at least 10-fold in response to chemical stress by aromatic aldehydes. Under the same growth conditions, similar enhanced in vivo levels of Sso2536 gene transcript were also measured. The gene encoding BldR was expressed in E. coli, and the recombinant protein was purified to homogeneity. DNA binding assays demonstrated that the protein is indeed a transcription factor able to recognize site specifically both the Sso2536 and mar-like promoters at sites containing palindromic consensus sequences. Benzaldehyde, the substrate of ADH(Ss), stimulates DNA binding of BldR at both promoters. The role of BldR in the auto-activation as well as in the regulation of the Sso2536 gene, together with results of increased operon and gene expression under conditions of exposure to aromatic aldehydes, indicates a novel coordinate regulatory mechanism in cell defense against stress by aromatic compounds.
在嗜热栖热菌(Sulfolobus solfataricus)中鉴定出一种DNA结合蛋白BldR,该蛋白在有毒醛类存在下生长的细胞中含量比正常细胞高5至10倍;它与醇脱氢酶基因(Sso2536)上游的调控序列结合。BldR与介导对多种环境应激反应的MarR(多重抗生素抗性)家族转录调节因子的细菌代表同源。转录分析表明,bldR基因在一个双顺反子单元中转录,该单元由编码转录调节因子(Sso1352)的基因和上游一个假定的多药转运蛋白(Sso1351)组成。通过与细菌对应物的同源性,这个双顺反子被命名为mar样操纵子。发现mar样操纵子的表达水平在芳香醛引起的化学应激下至少增加10倍。在相同的生长条件下,还检测到Sso2536基因转录本在体内水平有类似的增强。编码BldR的基因在大肠杆菌中表达,重组蛋白被纯化至同质。DNA结合试验表明,该蛋白确实是一种转录因子,能够在含有回文共有序列的位点特异性识别Sso2536和mar样启动子。ADH(Ss)的底物苯甲醛刺激BldR在两个启动子处的DNA结合。BldR在Sso2536基因的自激活以及调控中的作用,以及在暴露于芳香醛条件下操纵子和基因表达增加的结果,表明在细胞对芳香化合物应激的防御中存在一种新的协同调节机制。