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本文引用的文献

1
Subunit assembly in vivo of Escherichia coli RNA polymerase: role of the amino-terminal assembly domain of alpha subunit.大肠杆菌RNA聚合酶在体内的亚基组装：α亚基氨基末端组装结构域的作用
Genes Cells. 1996 Jun;1(6):517-28. doi: 10.1046/j.1365-2443.1996.d01-258.x.
2
Synthesis of the protein cutting reagent iron (S)-1-(p-bromoacetamidobenzyl)ethylenediaminetetraacetate and conjugation to cysteine side chains.蛋白质切割试剂铁（S）-1-（对溴乙酰氨基苄基）乙二胺四乙酸的合成及其与半胱氨酸侧链的偶联。
Bioconjug Chem. 1997 Jan-Feb;8(1):44-8. doi: 10.1021/bc9600731.
3
Upstream interactions at the lambda pRM promoter are sequence nonspecific and activate the promoter to a lesser extent than an introduced UP element of an rRNA promoter.λ pRM 启动子的上游相互作用不具有序列特异性，且与 rRNA 启动子引入的 UP 元件相比，对启动子的激活程度较低。
J Bacteriol. 1996 Dec;178(23):6945-51. doi: 10.1128/jb.178.23.6945-6951.1996.
4
Transcription factor recognition surface on the RNA polymerase alpha subunit is involved in contact with the DNA enhancer element.RNA聚合酶α亚基上的转录因子识别表面参与与DNA增强子元件的接触。
EMBO J. 1996 Aug 15;15(16):4358-67.
5
Ambidextrous transcriptional activation by SoxS: requirement for the C-terminal domain of the RNA polymerase alpha subunit in a subset of Escherichia coli superoxide-inducible genes.SoxS介导的双向转录激活：大肠杆菌超氧化物诱导基因亚群中RNA聚合酶α亚基C末端结构域的需求
Mol Microbiol. 1996 Jan;19(2):307-17. doi: 10.1046/j.1365-2958.1996.368893.x.
6
Determinants of RNA polymerase alpha subunit for interaction with beta, beta', and sigma subunits: hydroxyl-radical protein footprinting.RNA聚合酶α亚基与β、β'和σ亚基相互作用的决定因素：羟基自由基蛋白质足迹法
Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10162-6. doi: 10.1073/pnas.93.19.10162.
7
Identification of an UP element within the IHF binding site at the PL1-PL2 tandem promoter of bacteriophage lambda.在噬菌体λ的PL1-PL2串联启动子的整合宿主因子（IHF）结合位点内鉴定一个上游元件（UP元件）。
J Mol Biol. 1996 Jul 26;260(4):484-91. doi: 10.1006/jmbi.1996.0416.
8
Site-directed hydroxyl radical probing of the rRNA neighborhood of ribosomal protein S5.核糖体蛋白S5的rRNA邻域的定点羟基自由基探测
Science. 1996 Jun 14;272(5268):1659-62. doi: 10.1126/science.272.5268.1659.
9
Anatomy of a flexer-DNA complex inside a higher-order transposition intermediate.高阶转座中间体内部的Flexer-DNA复合物的结构剖析
Cell. 1996 May 31;85(5):761-71. doi: 10.1016/s0092-8674(00)81241-6.
10
Transcriptional activation of promoters of the superoxide and multiple antibiotic resistance regulons by Rob, a binding protein of the Escherichia coli origin of chromosomal replication.Rob（一种大肠杆菌染色体复制起点的结合蛋白）对超氧化物和多重抗生素抗性调节子启动子的转录激活作用。
J Bacteriol. 1996 May;178(9):2507-13. doi: 10.1128/jb.178.9.2507-2513.1996.

大肠杆菌RNA聚合酶的两个α亚基呈不对称排列，并与DNA上游元件的不同半区接触。

The two alpha subunits of Escherichia coli RNA polymerase are asymmetrically arranged and contact different halves of the DNA upstream element.

作者信息

Murakami K, Kimura M, Owens J T, Meares C F, Ishihama A

机构信息

Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.

出版信息

Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1709-14. doi: 10.1073/pnas.94.5.1709.

DOI:10.1073/pnas.94.5.1709
PMID:9050843
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC19981/
Abstract

RNA polymerase core enzyme of Escherichia coli is composed of two alpha subunits and one each of the beta and beta' subunits. The C-terminal domain of the RNA polymerase alpha subunit plays a key role in molecular communications with class I transcription factors and upstream (UP) elements of promoter DNA, using the same protein surface. To identify possible differences in the functional roles of the two alpha subunits, we have developed a reconstitution method for hybrid RNA polymerases containing two distinct alpha subunit derivatives in a defined orientation ("oriented alpha-heterodimer"). The binding sites of two alpha C-terminal domains on the UP element DNA were determined by hydroxyl radical-based DNA cleavage mediated by (p-bromoacetamidobenzyl)-EDTA x Fe, which was bound at Cys-269 on the UP recognition surface of one or both alpha subunits. The results clearly indicated that the two alpha subunits bind in tandem to two helix turns of the rrnBP1 UP element, and that the beta'-associated alpha subunit is bound to the promoter-distal region.

摘要

大肠杆菌的RNA聚合酶核心酶由两个α亚基以及各一个β和β'亚基组成。RNA聚合酶α亚基的C末端结构域在与I类转录因子和启动子DNA的上游（UP）元件进行分子通讯时发挥关键作用，且使用相同的蛋白质表面。为了确定两个α亚基在功能作用上可能存在的差异，我们开发了一种重组方法，用于构建在特定方向上含有两种不同α亚基衍生物的杂交RNA聚合酶（“定向α-异二聚体”）。通过由（对溴乙酰氨基苄基）-EDTA·Fe介导的基于羟基自由基的DNA切割，确定了两个α C末端结构域在UP元件DNA上的结合位点，该物质结合在一个或两个α亚基的UP识别表面的半胱氨酸-269位点上。结果清楚地表明，两个α亚基串联结合到rrnBP1 UP元件的两个螺旋圈上，并且与β'相关的α亚基结合到启动子远端区域。