Replication protein A in Pyrococcus furiosus is involved in homologous DNA recombination
Replication protein A in Pyrococcus furiosus is involved in homologous DNA recombination
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DOI：
10.1074/jbc.m102423200
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发表时间：
2001-07-13
影响因子：
4.8
通讯作者：
Ishino, Y
中科院分区：
文献类型：
--
作者：
Komori, K;Ishino, Y
细菌中的单链DNA结合蛋白和真核生物中的复制蛋白A（RPA）在DNA复制、修复和重组过程中起着至关重要的作用。我们从极端嗜热古菌Pyrococcus furiosus中鉴定出一种RPA复合物。与产甲烷古菌、詹氏甲烷球菌和热自养甲烷热杆菌的单肽RPA不同，强烈毕赤酵母RPA（PfuRPA）是由RPA 41、RPA 14和RPA 32三个亚基组成的稳定的异源寡聚复合物。RPA 41的氨基酸序列与真核生物RPA 70亚基和M.另一方面，RPA 14和RPA 32与细菌和真核生物的任何已知开放阅读框架都不具有同源性，然而，已公布的8种古细菌的全基因组序列中有6种具有与RPA 32同源的开放阅读框架。PfuRPA复合物，但不是每个单独的亚基，特异性地结合到单链DNA，并明显提高了体外链交换反应的效率，由P，furiosus RadA蛋白。此外，免疫沉淀分析表明，PfuRPA相互作用的重组蛋白，RadA和Hjc，以及复制蛋白，DNA聚合酶，引物酶，增殖细胞核抗原，复制因子C在P，furiosus细胞。这些结果表明，PfuRPA在P.
Single-stranded DNA-binding protein in Bacteria and replication protein A (RPA) in Eukarya play crucial roles in DNA replication, repair, and recombination processes. We identified an RPA complex from the hyperthermophilic archaeon, Pyrococcus furiosus. Unlike the single-peptide RPAs from the methanogenic archaea, Methanococcus jannaschii and Methanothermobacter thermoautotrophicus, P, furiosus RPA (PfuRPA) exists as a stable hetero-oligomeric complex consisting of three subunits, RPA41, RPA14, and RPA32, The amino acid sequence of RPA41 has some similarity to those of the eukaryotic RPA70 subunit and the M. jannaschii RPA, On the other hand, RPA14 and RPA32 do not share homology with any known open reading frames from Bacteria and Eukarya, However, six of eight archaea, whose total genome sequences have been published, have the open reading frame homologous to RPA32. The PfuRPA complex, but not each subunit alone, specifically bound to a single-stranded DNA and clearly enhanced the efficiency of an in vitro strand-exchange reaction by the P, furiosus RadA protein. Moreover, immunoprecipitation analyses showed that PfuRPA interacts with the recombination proteins, RadA and Hjc, as well as replication proteins, DNA polymerases, primase, proliferating cell nuclear antigen, and replication factor C in P, furiosus cells. These results indicate that PfuRPA plays important roles in the homologous DNA recombination in P. furiousus.