Replication Protein A in Pyrococcus furiosus is Involved in Homologous DNA Recombination

Kayoko Komori, Yoshizumi Ishino

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

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. furiosus.

Original languageEnglish
Pages (from-to)25654-25660
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number28
DOIs
Publication statusPublished - Jul 13 2001
Externally publishedYes

Fingerprint

Replication Protein A
Pyrococcus furiosus
Homologous Recombination
DNA
Archaea
Bacteria
Replication Protein C
Methanocaldococcus
DNA Primase
Proteins
Single-Stranded DNA
Proliferating Cell Nuclear Antigen
DNA-Binding Proteins
DNA-Directed DNA Polymerase
Eukaryota
Genetic Recombination
Open Reading Frames
Methanobacteriaceae
Repair
Genes

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Replication Protein A in Pyrococcus furiosus is Involved in Homologous DNA Recombination. / Komori, Kayoko; Ishino, Yoshizumi.

In: Journal of Biological Chemistry, Vol. 276, No. 28, 13.07.2001, p. 25654-25660.

Research output: Contribution to journalArticle

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