大肠杆菌RNA聚合酶依赖DNA的RNA合成：RNA链从DNA模板上的释放与重新起始
DNA-dependent synthesis of RNA by Escherichia coli RNA polymerase: release and reinitiation of RNA chains from DNA templates.
作者信息
Maitra U, Barash F
出版信息
Proc Natl Acad Sci U S A. 1969 Oct;64(2):779-86. doi: 10.1073/pnas.64.2.779.
Abstract
RNA synthesis in an in vitro RNA polymerase system at low ionic strength soon ceases, due to inhibition by accumulated RNA. Measurement of RNA chain initiation by the incorporation of gamma-(32)P-ATP and GTP with native T2 or T4 DNA as template shows that only one RNA chain is formed per molecule of enzyme added. In contrast, when the polymerase reaction is carried out in 10 mM Mg(++) and 0.2 M KCl, RNA synthesis proceeds nearly linearly for hours, resulting in a marked increase in accumulated RNA. Incorporation of gamma-(32)P-ATP also proceeds throughout the course of the reaction and the number of chains initiated per molecule of enzyme is increased severalfold. Most RNA chains formed are released from the DNA template as free RNA. Polymerase is released also in this process and acting catalytically reinitiates new chains. Rifampicin inhibits initiation of RNA synthesis and also blocks reinitiation of RNA chains without affecting growth of RNA chains already initiated.
摘要
在低离子强度的体外RNA聚合酶系统中，RNA合成很快就会停止，这是由于积累的RNA产生抑制作用。以天然T2或T4 DNA为模板，通过掺入γ-(32)P-ATP和GTP来测量RNA链起始，结果表明每添加一分子酶仅形成一条RNA链。相比之下，当聚合酶反应在10 mM Mg(++)和0.2 M KCl中进行时，RNA合成可持续数小时几乎呈线性进行，导致积累的RNA显著增加。γ-(32)P-ATP的掺入在整个反应过程中也持续进行，并且每分子酶起始的链数增加了几倍。形成的大多数RNA链作为游离RNA从DNA模板上释放出来。在此过程中聚合酶也被释放，并催化重新起始新的链。利福平抑制RNA合成的起始，并且还阻断RNA链的重新起始，而不影响已经起始的RNA链的生长。
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