The eukaryotic leading and lagging strand DNA polymerases are loaded onto primer-ends via separate mechanisms but have comparable processivity in the presence of PCNA.
The eukaryotic leading and lagging strand DNA polymerases are loaded onto primer-ends via separate mechanisms but have comparable processivity in the presence of PCNA.
复制标题
真核铅和滞后链DNA聚合酶通过单独的机制加载到引物上，但在PCNA存在下具有可比的加工性。
DOI：
10.1093/nar/gkm741
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发表时间：
2007
影响因子：
14.9
通讯作者：
Johansson E
中科院分区：
文献类型：
--
作者：
Chilkova O;Stenlund P;Isoz I;Stith CM;Grabowski P;Lundström EB;Burgers PM;Johansson E
酿酒酵母DNA聚合酶δ（Pol δ）和DNA聚合酶ε（Pol ε）是位于复制叉的复制型DNA聚合酶。这两种酶都受到PCNA的刺激，尽管水平不同。为了理解原因并探索与PCNA的相互作用，我们比较了Pol δ和Pol ε与PCNA和核酸（有或没有RPA）的物理相互作用，以及测量活性和持续合成能力的功能测定。利用表面等离子体共振技术，我们发现Pol ε对DNA具有高亲和力，但对PCNA具有低亲和力。相反，Pol δ对DNA具有低亲和力，对PCNA具有高亲和力。首次在RPA、PCNA和RFC存在的单链DNA上测量了Pol δ和Pol ε的真实持续合成能力。值得注意的是，在PCNA存在下，Pol δ和Pol ε对RPA包被的DNA的持续合成能力是相当的。最后，与Pol δ相比，Pol ε复制后的模板上有更多的PCNA分子。我们得出结论，Pol ε和Pol δ表现出相当的持续合成能力，但通过不同的机制加载在引物末端。
Saccharomyces cerevisiae DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) are replicative DNA polymerases at the replication fork. Both enzymes are stimulated by PCNA, although to different levels. To understand why and to explore the interaction with PCNA, we compared Pol δ and Pol ε in physical interactions with PCNA and nucleic acids (with or without RPA), and in functional assays measuring activity and processivity. Using surface plasmon resonance technique, we show that Pol ε has a high affinity for DNA, but a low affinity for PCNA. In contrast, Pol δ has a low affinity for DNA and a high affinity for PCNA. The true processivity of Pol δ and Pol ε was measured for the first time in the presence of RPA, PCNA and RFC on single-stranded DNA. Remarkably, in the presence of PCNA, the processivity of Pol δ and Pol ε on RPA-coated DNA is comparable. Finally, more PCNA molecules were found on the template after it was replicated by Pol ε when compared to Pol δ. We conclude that Pol ε and Pol δ exhibit comparable processivity, but are loaded on the primer-end via different mechanisms.
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影响因子：
5.3
作者：
Iida, T;Araki, H
通讯作者：
Araki, H
DOI：
10.1073/pnas.0505949102
发表时间：
2005-12-20
影响因子：
11.1
作者：
Garg, P;Burgers, PM
通讯作者：
Burgers, PM
影响因子：
5.6
作者：
Karthikeyan, R;Vonarx, EJ;Kunz, BA
通讯作者：
Kunz, BA
影响因子：
2.1
作者：
Fukui, T;Yamauchi, K;Waga, S
通讯作者：
Waga, S
影响因子：
4.8
作者：
Maki, S;Hashimoto, K;Sugino, A
通讯作者：
Sugino, A