Identification of the critical region in Replication factor C from Pyrococcus furiosus for the stable complex formation with Proliferating cell nuclear antigen and DNA

• Nishida Hirokazu

  Department of Structural Biology, Biomolecular Engineering Research Institute (BERI)

• Ishino Sonoko

  Department of Molecular Biology, Biomolecular Engineering Research Institute (BERI)

• Miyata Tomoko

  Department of Structural Biology, Biomolecular Engineering Research Institute (BERI)

• Morikawa Kosuke

  Department of Structural Biology, Biomolecular Engineering Research Institute (BERI)

• Ishino Yoshizumi

  Department of Molecular Biology, Biomolecular Engineering Research Institute (BERI) Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University

Abstract

Replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) are accessory proteins essential for processive DNA synthesis. The function of RFC is to load PCNA, a processivity factor of replicative DNA polymerases, onto primed DNA templates. The central hole of the PCNA homo-trimeric ring encircles doublestranded DNA, so that DNA polymerases can operate for DNA synthesis with PCNA along a DNA template. The Pyrococcus furiosus RFC (PfuRFC) consists of a small subunit (RFCS, 37kDa) and a large subunit (RFCL, 55kDa), which show significant sequence identity to the eukaryotic homologs. The C-terminal region of RFCL has an acidic cluster of about 30 amino acids, which consists mainly of glutamic acid residues, and a following basic cluster of 10 amino acids, which consists mainly of lysine residues. These clusters of charged amino acids, which precede the C-terminal consensus sequence, PIP (PCNA interacting protein)-box, are conserved in several archaeal RFCLs. The series of mutant PfuRFC containing the C-terminal deletions in RFCL were constructed. The mutational analyses showed that the charged cluster is not essential for loading of PCNA onto DNA. However, the region containing the basic cluster is important for the stable ternary (RFC-PCNA-DNA) complex formation.<br>

Journal

• Genes & Genetic Systems

  Genes & Genetic Systems 80 (2), 83-93, 2005

  The Genetics Society of Japan

Keywords

• archaea
• DNA replication
• proliferating cell nuclear antigen
• <i>Pyrococcus furiosus</i>
• replication factor c

Details

• CRID: 1390001205472519680
• NII Article ID: 130000062161
• NII Book ID: AA11077421
• DOI: 10.1266/ggs.80.83
• ISSN: 18805779
• ISSN: 13417568
• NDL BIB ID: 7346907
• PubMed: 16172520
• Web Site: http://id.ndl.go.jp/bib/7346907
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7346907
• Web Site: http://www.jstage.jst.go.jp/article/ggs/80/2/80_2_83/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• Abstract License Flag: Disallowed