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金属调控的mer启动子处,依赖激活因子的σ-70 RNA聚合酶预诱导结合。

Activator-dependent preinduction binding of sigma-70 RNA polymerase at the metal-regulated mer promoter.

作者信息

Heltzel A, Lee I W, Totis P A, Summers A O

机构信息

Department of Microbiology, University of Georgia, Athens 30602.

出版信息

Biochemistry. 1990 Oct 16;29(41):9572-84. doi: 10.1021/bi00493a011.

Abstract

Expression of the Tn21 mercury-resistance (mer) locus is controlled by the merR gene product, which represses mer structural gene (merTPCAD) transcription in the absence of mercuric ion [Hg(II)] and activates it in the presence of Hg(II). In vivo DNA methylation of the mer regulatory region (merOP) shows that, with or without the inducer Hg(II), MerR strongly protects four guanine residues in a dyadic region located between the -10 and -35 hexamers of the structural gene promoter (PTPCAD). Prior to induction by Hg(II), RNA polymerase is also bound at PTPCAD; occupancy of the uninduced promoter by RNA polymerase is dependent on MerR. Methylation and permanganate footprinting demonstrate that induction by Hg(II) results in MerR/Hg(II)-dependent promoter DNA melting in the -10 region of PTPCAD and in additional DNA structural distortions within the region of dyad symmetry. Thus, MerR fosters the binding of RNA polymerase to an inactive promoter, and upon induction, MerR/Hg(II) facilitates DNA distortions suitable for efficient formation of the active transcription complex.

摘要

Tn21汞抗性(mer)基因座的表达受merR基因产物控制,在没有汞离子[Hg(II)]的情况下,merR基因产物会抑制mer结构基因(merTPCAD)的转录,而在有Hg(II)存在时则会激活转录。mer调控区(merOP)的体内DNA甲基化显示,无论有无诱导剂Hg(II),MerR都能强烈保护结构基因启动子(PTPCAD)的-10和-35六聚体之间二元区域中的四个鸟嘌呤残基。在被Hg(II)诱导之前,RNA聚合酶也结合在PTPCAD上;RNA聚合酶对未诱导启动子的占据依赖于MerR。甲基化和高锰酸盐足迹分析表明,Hg(II)诱导会导致PTPCAD的-10区域中MerR/Hg(II)依赖性的启动子DNA解链,并在二元对称区域内产生额外的DNA结构畸变。因此,MerR促进RNA聚合酶与无活性启动子的结合,诱导后,MerR/Hg(II)促进适合高效形成活性转录复合物的DNA畸变。

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