Comparative studies of the effect of DNA superhelicity on in vitro transcription catalyzed by Escherichia coli S100 proteins and purified RNA polymerase.
Comparative studies of the effect of DNA superhelicity on in vitro transcription catalyzed by Escherichia coli S100 proteins and purified RNA polymerase.
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DNA超螺旋对大肠杆菌S100蛋白和纯化RNA聚合酶催化体外转录影响的比较研究。
DOI：
10.1016/0378-1119(82)90071-3
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发表时间：
1982
期刊：
影响因子：
3.5
通讯作者：
F. Imamoto
中科院分区：
文献类型：
--
作者：
K. Kuroki;S. Ishii;Y. Kano;F. Imamoto
在不同的Kc1浓度下，研究了DNA超螺旋对纯化的大肠杆菌RNA聚合酶或S100粗提物蛋白体外转录的影响。以含有pL启动子控制的N-trp基因和pR启动子控制的cro基因的重组质粒pMT48的DNA为模板。与纯RNA聚合酶相比，S100粗提物蛋白对转录的刺激通常更明显。在Kc1浓度低于100 mM的纯RNA聚合酶作用下，pMT48DNA的模板活性在超螺旋形式和松弛形式之间没有显著差异。相反，在使用S100粗提物的体系中，在从1.7到400 mM的整个KCl浓度范围内，有效模板活性对超螺旋度的依赖性很大。在这两种转录检测系统中都可以观察到pR启动子对超螺旋的相对不敏感。对pL促进的trpmRNA合成的动力学分析表明，在松弛的模板上体外转录减少的主要原因是RNA链的启动频率较低，但至少部分原因是链延长的过早停止。
The effect of DNA superhelicity on in vitro transcription catalyzed by purifiedEscherichia coliRNA polymerase or S100 crude extract proteins was examined at various KC1 concentrations. DNA from a recombinant plasmid pMT48 harboring thepLpromoter-controlled fusedN-trpgenes and thepRpromoter-controlledtof(cro) gene was employed as a template. Stimulation of transcription by superhelicity is generally more pronounced with the S100 crude extract proteins than with pure RNA polymerase. At KC1 concentrations lower than 100 mM with pure RNA polymerase, there is no significant difference in the template activity between the supercoiled and relaxed forms of pMT48 DNA. In contrast, the dependence of efficient template activity on superhelicity is great over a whole range of KCl concentrations from 1.7 to 400 mM in the system using the S100 crude extract. The relative insensitivity of thepRpromoter to superhelicity can be observed in either transcription assay system. Analysis of the kinetics ofpL-promoted synthesis oftrpmRNA indicates that diminished transcription in vitro on a relaxed template results mainly from less frequent RNA chain initiations, but at least in part from premature arrest of the chain elongation.
DNA 结构和基因调控。
DOI：
10.1016/s0079-6603(08)60674-1
发表时间：
1980
期刊：
Progress in nucleic acid research and molecular biology
影响因子：
--
作者：
Wells,RD;Goodman,TC;Hillen,W;Horn,GT;Klein,RD;Larson,JE;Muller,UR;Neuendorf,SK;Panayotatos,N;Stirdivant,SM
通讯作者：
Stirdivant,SM