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首页> 外文期刊>Journal of Molecular Biology >Processive proofreading and the spatial relationship between polymerase and exonuclease active sites of bacteriophage phi29 DNA polymerase.
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Processive proofreading and the spatial relationship between polymerase and exonuclease active sites of bacteriophage phi29 DNA polymerase.

机译：噬菌体phi29 DNA聚合酶的聚合校对和聚合酶与核酸外切酶活性位点之间的空间关系。

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摘要

phi29 DNA polymerase is a multifunctional enzyme, able to incorporate and to proofread misinserted nucleotides, maintaining a very high replication fidelity. Since both activities are functionally separated, a mechanism is needed to guarantee proper coordination between synthesis and degradation, implying movement of the DNA primer terminus between polymerization and 3'-5' exonuclease active sites. Using single-turnover conditions, we have demonstrated that phi29 DNA polymerase edits the polymerization errors using an intramolecular pathway; that is, the primer terminus travels from one active site to the other without dissociation from the DNA. On the other hand, by using chemical tags, we could infer a difference in length of only one nucleotide to contact the primer strand when it is in the polymerization mode versus the editing mode. Using the same approach, it was estimated that phi29 DNA polymerase covers a DNA region of ten nucleotides, as has been measured in other polymerases using different techniques. Copyright 1999 Academic Press.
机译：phi29 DNA聚合酶是一种多功能酶，能够整合并校对错误插入的核苷酸，从而保持很高的复制保真度。由于两种活性在功能上是分开的，因此需要一种机制来保证合成和降解之间的适当协调，这意味着DNA引物末端在聚合和3'-5'核酸外切酶活性位点之间运动。使用单周转条件，我们已经证明phi29 DNA聚合酶使用分子内途径来编辑聚合错误。也就是说，引物末端从一个活性位点移动到另一个活性位点，而没有与DNA分离。另一方面，通过使用化学标签，我们可以推断出在聚合模式与编辑模式下，仅一个核苷酸接触引物链的长度差异。使用相同的方法，估计phi29 DNA聚合酶覆盖了十个核苷酸的DNA区域，正如其他聚合酶中使用不同技术所测量的那样。版权所有1999，学术出版社。

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