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首页> 外文期刊>Journal of Molecular Biology >Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B.
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Binding to PCNA in Euryarchaeal DNA Replication requires two PIP motifs for DNA polymerase D and one PIP motif for DNA polymerase B.

机译：与Euryarchaeal DNA复制中的PCNA结合需要DNA聚合酶D的两个PIP基序和DNA聚合酶B的一个PIP基序。

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摘要

Replicative DNA polymerases possess a canonical C-terminal proliferating cell nuclear antigen (PCNA)-binding motif termed the PCNA-interacting protein (PIP) box. We investigated the role of the PIP box on the functional interactions of the two DNA polymerases, PabPol B (family B) and PabPol D (family D), from the hyperthermophilic euryarchaeon Pyrococcus abyssi, with its cognate PCNA. The PIP box was essential for interactions of PabPol B with PCNA, as shown by surface plasmon resonance and primer extension studies. In contrast, binding of PabPol D to PCNA was affected only partially by removing the PIP motif. We identified a second palindromic PIP box motif at the N-terminus of the large subunit of PabPol D that was required for the interactions of PabPol D with PCNA. Thus, two PIP motifs were needed for PabPol D for binding to PabPCNA. Moreover, the C-terminus of PabPCNA was essential for stimulation of PabPol D activity but not for stimulation of PabPol B activity. Neither DNA polymerase interacted with the PabPCNA interdomain connecting loop. Our data suggest that distinct processes are involved in PabPol D and PabPol B binding to PCNA, raising the possibility that Archaea require two mechanisms for recruiting replicative DNA polymerases at the replication fork.
机译：复制性DNA聚合酶具有典型的C端增殖细胞核抗原（PCNA）结合基序，称为PCNA相互作用蛋白（PIP）盒。我们调查了PIP盒在来自超嗜热性球菌毕赤酵母的两个DNA聚合酶PabPol B（家族B）和PabPol D（家族D）与其相关的PCNA的功能相互作用中的作用。如表面等离子体共振和引物延伸研究所示，PIP盒对于PabPol B与PCNA的相互作用至关重要。相反，通过去除PIP基序，PabPol D与PCNA的结合仅部分受到影响。我们在PabPol D与PCNA的相互作用所必需的PabPol D大亚基的N末端确定了第二个回文PIP盒基序。因此，PabPol D需要两个PIP基序才能与PabPCNA结合。此外，PabPCNA的C末端对于刺激PabPol D活性是必不可少的，但对于刺激PabPol B活性却不是必需的。两种DNA聚合酶均未与PabPCNA域间连接环相互作用。我们的数据表明PabPol D和PabPol B与PCNA的结合涉及不同的过程，这增加了古细菌需要两种机制在复制叉处募集复制性DNA聚合酶的可能性。

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