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Coupling of transcription and replication machineries in λ DNA replication initiation: evidence for direct interaction of Escherichia coli RNA polymerase and the λO protein

机译:转录和复制机制在λDNA复制起始中的耦合:大肠杆菌RNA聚合酶与λO蛋白直接相互作用的证据

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摘要

Transcription proceeding downstream of the λ phage replication origin was previously shown to support initial steps of the λ primosome assembly in vitro and to regulate frequency and directionality of λ DNA replication in vivo. In this report, the data are presented indicating that the RNA polymerase β subunit makes a direct contact with the λO protein, a replication initiator of λ phage. These results suggest that the role of RNA polymerase during the initiation of λ phage DNA replication may be more complex than solely influencing DNA topology. Results demonstrated in this study also show that gyrase supercoiling activity stimulates the formation of a complex between λO and RNA polymerase, suggesting that the introduction of negative supercoils by DNA gyrase, besides lowering the energy required for DNA strand separation, may play an additional role in modeling protein–protein interactions at early steps of DNA replication initiation.
机译:先前已经证明,在λ噬菌体复制起点下游进行的转录支持体外的λprimosome装配的初始步骤,并在体内调节λDNA复制的频率和方向。在此报告中,提供的数据表明RNA聚合酶β亚基与λO蛋白(λ噬菌体的复制引发剂)直接接触。这些结果表明,RNA聚合酶在λ噬菌体DNA复制起始过程中的作用可能比仅影响DNA拓扑结构更为复杂。这项研究证明的结果还表明,回旋酶的超螺旋活性刺激了λO和RNA聚合酶之间的复合物的形成,这表明DNA回旋酶引入负性超螺旋,除了降低了DNA链分离所需的能量外,还可能在在DNA复制起始的早期阶段模拟蛋白质之间的相互作用。

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