DNA与RNA聚合酶相互作用的物理化学研究。大肠杆菌RNA聚合酶使DNA螺旋解旋。
Physiochemical studies on interactions between DNA and RNA polymerase. Unwinding of the DNA helix by Escherichia coli RNA polymerase.
作者信息
Wang J C, Jacobsen J H, Saucier J M
出版信息
Nucleic Acids Res. 1977;4(5):1225-41. doi: 10.1093/nar/4.5.1225.
Abstract
In a medium containing 10mM Tris, pH 8, 10 mM MG++, 50 mM K+ and 10 mM NH4, the binding of an E. coli RNA polymerase holoenzyme unwinds the DNA helix by about 240 degrees at 37 degrees C. In this medium the total unwinding of the DNA increases linearly with the molar ratio of polymerase to DNA. The number of binding sites at which unwinding can occur is very large. If the K+ concentration is increased at 200 mM, the enzyme binds to only a limited number of sites, and the bound and free enzyme molecules do not exchange at an appreciable rate. The unwinding angle of the DNA per bound enzyme in this high salt medium is measured to be 140 degrees at 37 degrees C. The total unwinding angle for a fixed number of bound polymerase molecules per DNA is strongly temperature dependent, and decreases with decreasing temperature.
摘要
在含有10mM Tris（pH 8）、10mM Mg++、50mM K+和10mM NH4的介质中，大肠杆菌RNA聚合酶全酶的结合在37℃时使DNA螺旋解开约240度。在这种介质中，DNA的总解旋随着聚合酶与DNA的摩尔比呈线性增加。能够发生解旋的结合位点数量非常多。如果将K+浓度增加到200mM，酶仅结合到有限数量的位点，并且结合态和游离态的酶分子不会以可观的速率交换。在这种高盐介质中，每个结合的酶在37℃时使DNA的解旋角度测得为140度。每个DNA上固定数量的结合聚合酶分子的总解旋角度强烈依赖于温度，并且随着温度降低而减小。
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