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Reconstitution of a eukaryotic replisome reveals the mechanism of asymmetric distribution of DNA polymerases

机译:真核复制体的重组揭示了DNA聚合酶不对称分布的机制

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摘要

Eukaryotes require 3 DNA polymerases for normal replisome operations, DNA polymerases (Pol) alpha, delta and epsilon. Recent biochemical and structural studies support the asymmetric use of these polymerases on the leading and lagging strands. Pol epsilon interacts with the 11-subunit CMG helicase, forming a 15-protein leading strand complex that acts processively in leading strand synthesis in vitro, but Pol epsilon is inactive on the lagging strand. The opposite results are observed for Pol delta with CMG. Pol delta is highly active on the lagging strand in vitro, but has only feeble activity with CMG on the leading strand. Pol a also functions with CMG to prime both strands, and is even capable of extending both strands with CMG present. However, extensive DNA synthesis by Pol a is sharply curtailed by the presence of either Pol epsilon or Pol delta, which limits the role of the low fidelity Pol a to the initial priming of synthesis.
机译:真核生物需要3种DNA聚合酶才能进行正常的复制操作,即DNA聚合酶(Pol)alpha,δ和epsilon。最近的生化和结构研究支持这些聚合酶在前导链和滞后链上的不对称使用。 Pol epsilon与11个亚基CMG解旋酶相互作用,形成15蛋白前导链复合物,该复合物在体外前导链合成中起着过程性作用,但Pol epsilon在落后链上无活性。对于使用CMG的Pol delta观察到相反的结果。波尔塔三角洲在体外对滞后链具有很高的活性,但对前链的CMG仅具有微弱的活性。 Pol a还可与CMG一起启动两条链,甚至能够在存在CMG的情况下延伸两条链。但是,Pol a的广泛DNA合成由于Pol epsilon或Pol delta的存在而大大减少,这将低保真度Pol a的作用限制于合成的初始引发。

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