Saridakis Vivian, Shahinas Dea, Xu Xiaohui, Christendat Dinesh
Department of Biology, York University, 4700 Keele Street, Toronto, Ontario, Canada.
J Mol Biol. 2008 Mar 28;377(3):655-67. doi: 10.1016/j.jmb.2008.01.001. Epub 2008 Jan 11.
Transcriptional regulators belonging to the MarR family are characterized by a winged-helix DNA binding domain. These transcriptional regulators regulate the efflux and influx of phenolic agents in bacteria and archaea. In Escherichia coli, MarR regulates the multiple antibiotic resistance operon and its inactivation produces a multiple antibiotic resistance phenotype. In some organisms, active efflux of drug compounds will produce a drug resistance phenotype, whereas in other organisms, active influx of chlorinated hydrocarbons results in their rapid degradation. Although proteins in the MarR family are regulators of important biological processes, their mechanism of action is not well understood and structural information about how phenolic agents regulate the activity of these proteins is lacking. This article presents the three-dimensional structure of a protein of the MarR family, MTH313, in its apo form and in complex with salicylate, a known inactivator. A comparison of these two structures indicates that the mechanism of regulation involves a large conformational change in the DNA binding lobe. Electrophoretic mobility shift assay and biophysical analyses further suggest that salicylate inactivates MTH313 and prevents it from binding to its promoter region.
属于MarR家族的转录调节因子的特征是具有一个带翼螺旋DNA结合结构域。这些转录调节因子调控细菌和古细菌中酚类物质的外排和内流。在大肠杆菌中,MarR调节多重耐药操纵子,其失活会产生多重耐药表型。在一些生物体中,药物化合物的主动外排会产生耐药表型,而在其他生物体中,氯代烃的主动内流会导致它们快速降解。尽管MarR家族的蛋白质是重要生物学过程的调节因子,但其作用机制尚未完全了解,并且缺乏关于酚类物质如何调节这些蛋白质活性的结构信息。本文展示了MarR家族一种蛋白质MTH313的三维结构,包括其无配体形式以及与已知失活剂水杨酸形成的复合物形式。对这两种结构的比较表明,调节机制涉及DNA结合叶的巨大构象变化。电泳迁移率变动分析和生物物理分析进一步表明,水杨酸使MTH313失活并阻止其与启动子区域结合。