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首页> 外文期刊>Journal of Molecular Biology >ADJACENT UPSTREAM SUPERHELICAL WRITHE INFLUENCES AN ESCHERICHIA COLI PROMOTER AS MEASURED BY IN VIVO STRENGTH AND IN VITRO OPEN COMPLEX FORMATION
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ADJACENT UPSTREAM SUPERHELICAL WRITHE INFLUENCES AN ESCHERICHIA COLI PROMOTER AS MEASURED BY IN VIVO STRENGTH AND IN VITRO OPEN COMPLEX FORMATION

机译：邻近的上层超螺旋缠绕物对体内大肠杆菌强度和体外开放复合物形成的肠埃希氏菌启动子的影响

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摘要

This work investigates the effect on transcription of superhelical writhe located in the region immediately upstream of the -35 consensus sequence of Escherichia coli promoters. A set of double-stranded oligonucleotides, each with an unique DNA configuration, were designed, synthesized and substituted into an area of naturally occurring right-handed superhelical curvature immediately. upstream of the beta-lactamase promoter in plasmid pUC19. All the mutants showed reduced promoter activities in E. coli cells. However, rightward superhelical writhe clearly facilitated transcription when compared with the effect produced by a straight DNA segment. Leftward writhe greatly repressed transcription. A plane curve hewed an intermediate effect. This phenomenon was due not only to the difference in the ability of the segment to drive an open complex formation, but also to the difference in the binding affinity of RNA polymerase to the promoter. The positive effect of rightward writhe was also observed in vivo for the promoter of the tetracycline resistance gene of pBR322. The sense and extent of superhelical writhe of a DNA curvature seem to determine its influence on promoter activity. (C) 1995 Academic Press Limited [References: 42]
机译：这项工作研究了位于大肠杆菌启动子-35共有序列上游紧邻区域的超螺旋病毒对转录的影响。设计，合成了一组各自具有独特DNA构型的双链寡核苷酸，并将其立即替换为自然产生的右手超螺旋弯曲区域。质粒pUC19中β-内酰胺酶启动子的上游。所有突变体在大肠杆菌细胞中均显示启动子活性降低。然而，与直DNA片段产生的作用相比，右旋超螺旋明显促进了转录。左旋大大抑制了转录。平面曲线产生了中间效果。该现象不仅是由于片段驱动开放复合物形成的能力不同，还归因于RNA聚合酶与启动子的结合亲和力的不同。在体内也观察到了pBR322的四环素抗性基因的启动子向右旋转的积极作用。 DNA弯曲的超螺旋扭曲感和程度似乎决定了其对启动子活性的影响。 （C）1995 Academic Press Limited [参考：42]

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