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首页> 外文期刊>Journal of Molecular Biology >DNA polymerase switching: II. Replication factor C abrogates primer synthesis by DNA polymerase alpha at a critical length.
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DNA polymerase switching: II. Replication factor C abrogates primer synthesis by DNA polymerase alpha at a critical length.

机译：DNA聚合酶转换：II。复制因子C取消了DNA聚合酶α临界长度的引物合成。

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摘要

A crucial event in DNA replication is the polymerase switch from the synthesis of a short RNA/DNA primer by DNA polymerase alpha/primase to the pro?cessive elongation by DNA polymerase delta. In order to shed light on the role of replication factor C (RF-C) in this process, the effects of RF-C on DNA polymerase alpha were investigated. We show that RF-C stalls DNA polymerase alpha after synthesis of approximately 30 nucleotides, while not inhibiting the polymerase activity per se. This suggested that RF-C and the length of the primer may be two important factors contributing to the polymerase switch. Furthermore the DNA binding properties of RF-C were tested. Band shift experiments indicated that RF-C has a preference for 5' recessed ends and double-stranded DNA over 3' ends. Finally PCNA can be loaded onto a DNA template carrying a RNA primer, suggesting that a DNA moiety is not necessarily required for the loading of the clamp. Thus we propose a model where RF-C, upon binding to the RNA/DNA primer, influences primer synthesis and sets the conditions for a polymerase switch after recruiting PCNA to DNA. Copyright 2000 Academic Press.
机译：DNA复制中的关键事件是聚合酶从DNA聚合酶alpha / primase合成短RNA / DNA引物到DNA聚合酶delta逐步延伸。为了阐明复制因子C（RF-C）在此过程中的作用，研究了RF-C对DNA聚合酶α的影响。我们显示，RF-C合成大约30个核苷酸后会停滞DNA聚合酶α，而不会抑制聚合酶本身的活性。这表明RF-C和引物的长度可能是促成聚合酶转换的两个重要因素。此外，测试了RF-C的DNA结合特性。带移实验表明，RF-C偏爱5'的凹端和双链DNA优于3'端。最后，可以将PCNA装载到带有RNA引物的DNA模板上，这表明装载夹具不一定需要DNA部分。因此，我们提出了一个模型，其中RF-C与RNA / DNA引物结合后，会影响引物的合成，并为PCNA募集到DNA后设定了聚合酶转换的条件。版权所有2000学术出版社。

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