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大肠杆菌RNA聚合酶与λPR启动子之间形成开放复合物所需的DNA功能基团。通过碱基类似物取代进行鉴定。

DNA functional groups required for formation of open complexes between Escherichia coli RNA polymerase and the lambda PR promoter. Identification via base analog substitutions.

作者信息

Dubendorff J W, deHaseth P L, Rosendahl M S, Caruthers M H

出版信息

J Biol Chem. 1987 Jan 15;262(2):892-8.

PMID:2948952
Abstract

Synthetic 75-base pair promoters bearing base changes and/or base analog substitutions at selected positions were constructed. Using both abortive initiation and run-off transcription assays, the interaction of these altered promoters with Escherichia coli RNA polymerase was studied in order to determine the involvement of DNA functional groups in promoter recognition. Two adjacent thymines in the -35 region were identified whose 5-methyl groups play a crucial role. Additionally, the combined results from several substitution experiments showed that functional groups in the major groove of the strongly conserved T-A base pair at the -7 position are probable sites of direct interaction with RNA polymerase.

摘要

构建了在选定位置带有碱基变化和/或碱基类似物取代的合成75碱基对启动子。使用流产起始和延伸转录测定法，研究了这些改变的启动子与大肠杆菌RNA聚合酶的相互作用，以确定DNA功能基团在启动子识别中的作用。在-35区域鉴定出两个相邻的胸腺嘧啶，其5-甲基基团起着关键作用。此外，几个取代实验的综合结果表明，-7位高度保守的T-A碱基对大沟中的功能基团可能是与RNA聚合酶直接相互作用的位点。

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J Biol Chem. 1987 Jan 15;262(2):892-8.
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