Spassky A, Rimsky S, Buc H, Busby S
Institut Pasteur, Unité de Physicochimie des Macromolécules Biologiques, Paris, France.
EMBO J. 1988 Jun;7(6):1871-9. doi: 10.1002/j.1460-2075.1988.tb03020.x.
The lac and gal control regions contain two functional overlapping promoters P1 and P2. Point mutations can shift transcription from P1 to P2 and vice versa. We show that the reactivity of DNA fragments towards nucleolytic attack with orthophenanthroline cuprous complex can be used to predict which promoter competes more efficiently for RNA polymerase binding. Furthermore, similar changes in reactivity are observed as closed complexes isomerize to form the final open complexes, provided that the functional start is taken as a reference. We found a correlation between the reactivity pattern of -10 regions in uncomplexed DNA and the rate of open complex formation.
乳糖操纵子和半乳糖操纵子控制区域包含两个功能性重叠启动子P1和P2。点突变可使转录从P1转换至P2,反之亦然。我们表明,邻菲罗啉亚铜络合物对DNA片段的核酸酶攻击反应性可用于预测哪个启动子在与RNA聚合酶结合方面竞争更有效。此外,以功能起始位点为参考,当封闭复合物异构化为最终开放复合物时,会观察到类似的反应性变化。我们发现未复合DNA中-10区域的反应性模式与开放复合物形成速率之间存在相关性。