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Researches on functional link between DNA replication and chromosomal cohesion through PCNA clamp

Research Project

Project/Area Number 17370064
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionKyushu University

Principal Investigator

TSURIMOTO Toshiki  Kyushu University, faculty of Sciences, Professor (30163885)

Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥15,660,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2007: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2006: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 2005: ¥6,000,000 (Direct Cost: ¥6,000,000)
Keywordsreplication fork / replication apparatus / chromosomal cohesion / clamp / clamp loader / DNA polymerase
Research Abstract

1. Functional interaction of clamp loader complexes with DNA polymerase eta
One of clamp loader complexes, Ctf18-RFC, which consists of the chromosome cohesion factors Ctf18, Dccl, and Ctf8 and four small subunits of RFC, functions as a second PCNA loader. We identified Pol eta as a DNA polymerase activity stimulated by Ctf18-RFC but not by RFC. Ctf18-RFC stimulated Pol eta on its own, and the addition of PCNA cooperatively increased that stimulation. Furthermore, Ctf18-RFC interacted physically with pol eta. In addition, we observed the similar physical interaction of RFC with pol eta, but affecting its DNA synthesis activity differently from Ctf18-RFC. We propose that the novel loader-DNA polymerase interactions can switch the activity of a DNA replication fork to adapt to various template structures.
2. Molecular interaction between loader complexes and replication fork components
Using human cell lines ectopically expressing tagged loader complexes, FLAG-RFC, FLAG-(Rad17-RFC), FLAG-(Ctf18-RFC), we have identified Pol epsilon and RPA as common loader binding proteins. Further studies with recombinant proteins have indicated that their interactions are direct and specific. All loaders can associate with these proteins through their common small subunits, and also through specific subunits, suggesting roles to maintain functional protein assemblies at the replication fork.
3. Functions of WRNIP1 protein
We purified one of RFC-related proteins, WRNIP1, which has been identified as a Werner DNA helicase binding protein. This protein forms a self-oligomerized complex and exhibits ATPase activity. It also interacts specifically with replicative DNA polymerase delta and stimulates the activity, suggesting its role in regulation of DNA synthesis by Pol delta.

Report

(4 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (34 results)

All 2008 2007 2006 2005 Other

All Journal Article (19 results) (of which Peer Reviewed: 4 results) Presentation (6 results) Book (8 results) Remarks (1 results)

  • [Journal Article] DNA damage induced ubiquitylation of RFC2 subunit of RFC complex2008

    • Author(s)
      Tomida, J, et. al.
    • Journal Title

      J Biol Chem 283

      Pages: 9071-9079

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Recognition of forked and single-stranded DNA structures by human RAD18 complexed with RAD6Bprotein triggers its recruitment to stalled replication forks2008

    • Author(s)
      Tsuji, Y, et. al.
    • Journal Title

      Genes Cells 13

      Pages: 343-354

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] DNA damage induced ubiquitylation of RFC2 subunit of RFC complex2008

    • Author(s)
      Tomida, J.
    • Journal Title

      J Biol Chem 283

      Pages: 9071-9079

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Recognition of forked and single-stranded DNA structures by human RAD18 complexed with RAD6B protein triggers its recruitment to stalled replication forks2008

    • Author(s)
      Tsuji, Y.
    • Journal Title

      Genes Cells 13

      Pages: 343-354

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] DNA damage induced ubiquitylation of RFC2 subunit of RFC complex.2008

    • Author(s)
      Tomida, J., et. al.
    • Journal Title

      J Biol Chem (in press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Recognition of forked and single-stranded DNA structures by human RAD18 complexed with RAD6Bprotein triggers its recruitment to stalled replication forks.2008

    • Author(s)
      Tsuji, Y., et. al.
    • Journal Title

      Genes Cells (in press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A second PCNA loader complex, Ctf18-RFC, stimulates DNA polymerase h activity2007

    • Author(s)
      Shiomi, Y, et. al.
    • Journal Title

      J. Biol. Chem 282

      Pages: 20906-20914

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Dynamics of human replication relication factors in the elongation phase of DNA replication2007

    • Author(s)
      Masuda Y, et. al.
    • Journal Title

      Nucleic Acids Res 35

      Pages: 6904-6916

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, targethuman Cdt1 for proteolysis2007

    • Author(s)
      Nishitani H
    • Journal Title

      EMBO J 25

      Pages: 1126-1136

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Dynamics of human replication factors in the elongation phase of DNA replication2007

    • Author(s)
      Masuda, Y.
    • Journal Title

      Nucleic Acids Res 35

      Pages: 6904-6916

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] A second PCNA loader complex,Ctf18-RFC,stimulates DNA polymerase h activity2007

    • Author(s)
      Shiomi, Y., et. al.
    • Journal Title

      J.Biol.Chem. 282

      Pages: 20906-20914

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Dynamics of human replication factors in the elongation phase of DNA replication2007

    • Author(s)
      Masuda Y., et. al.
    • Journal Title

      Nucleic Acids Res. 35

      Pages: 6904-6916

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] PCNA and RFC proteins in Genomic Stability2006

    • Author(s)
      Tsurimoto, T.
    • Journal Title

      in DNA Replication and Human Disease. ed.DePamphilis, M. L, Cold Spring Harbor Press NY

      Pages: 411-434

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cu14, target human Cdt1 for proteolysis2006

    • Author(s)
      Nishitani, H.
    • Journal Title

      EMBO J'25

      Pages: 1126-1136

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cu14, target human Cdt1 for proteolysis.2006

    • Author(s)
      Nishitani H
    • Journal Title

      EMBO J 25

      Pages: 1126-1136

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Human Werner helicase interacting protein 1 (WRNIP1)functions as a novel modulator for DNA polymerase d2005

    • Author(s)
      Tsurimoto, T.
    • Journal Title

      Genes Cells 10

      Pages: 13-22

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Human Werner helicase interacting protein 1(WRNIP1) functions as a novel modulator for DNA polymerase delta.2005

    • Author(s)
      Tsurimoto, T.
    • Journal Title

      Genes Cells 10

      Pages: 13-22

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Human Werner helicase interacting protein 1(WRNIP1) functions as a novel modulator for DNA polymerase δ2005

    • Author(s)
      Tsurimoto, T.
    • Journal Title

      Genes Cells 10

      Pages: 13-22

    • Related Report
      2005 Annual Research Report
  • [Journal Article] PCNA-interacting proteins

    • Author(s)
      Tsurimoto, T.
    • Journal Title

      in Proliferating Cell Nuclear Antigen(PCNA). ed. Lee H. Research Signpost

      Pages: 25-49

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ローダー複合体とPolε、RPA間の相互作用の解析2008

    • Author(s)
      村上 武司
    • Organizer
      第25回染色体ワークショップ
    • Place of Presentation
      静岡県熱海市
    • Related Report
      2007 Annual Research Report
  • [Presentation] Functional interaction of clamp loader complexes with DNA polymeraseeta.2008

    • Author(s)
      Tsurimoto, T.
    • Organizer
      Eukaryotic DNA Replication & Genome Maintenance
    • Place of Presentation
      Cold Spring Harbor Lab.NY, USA
    • Related Report
      2007 Annual Research Report
  • [Presentation] Regulation of DNA polymerase eta activity by Clamp Loader Complexes.2007

    • Author(s)
      Tsurimoto, T.
    • Organizer
      elMBL Workshop on DNA replication
    • Place of Presentation
      Tokyo University, Japan
    • Year and Date
      2007-04-16
    • Related Report
      2007 Annual Research Report
  • [Presentation] クランプローダーとDNAポリメラーゼε間の相互作用の解析2007

    • Author(s)
      村上 武司
    • Organizer
      組換え・複製合同ワークショップ
    • Place of Presentation
      静岡県伊豆市
    • Related Report
      2007 Annual Research Report
  • [Presentation] クランプローダーとDNA複製因子の相互作用の解析2007

    • Author(s)
      村上 武司
    • Organizer
      日本分子生物学・生化学会合同年会
    • Place of Presentation
      神奈川県横浜市
    • Related Report
      2007 Annual Research Report
  • [Presentation] コビキチン化PCNAによるDNA polymeraseの活性促進2007

    • Author(s)
      廣川 雅人
    • Organizer
      日本分子生物学・生化学会合同年会
    • Place of Presentation
      神奈川県横浜市
    • Related Report
      2007 Annual Research Report
  • [Book] 複製フォークの舵取り役、クランプとクランプローダータンパク質(実験医学、25巻5号 増刊「染色体サイクル」)2007

    • Author(s)
      釣本 敏樹
    • Total Pages
      7
    • Publisher
      羊土社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report
  • [Book] 複製フォークの舵取り役、クランプとクランプローダータンパク質(実験医学、25巻5号増刊「染色体サイクル」)2007

    • Author(s)
      釣本 敏樹
    • Total Pages
      7
    • Publisher
      羊土社
    • Related Report
      2007 Annual Research Report
  • [Book] PCNA-interacting proteins(in Proliferating Cell Nuclear Antigen(PCNA)ed.Lee H.)2006

    • Author(s)
      Tsurimoto T
    • Total Pages
      25
    • Publisher
      Research Signpost, Kerala, India
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] PCNA Replication and RFC proteins in Genomic Stability.(in DNA replication and human disease.ed.DePamphilis, M.L, )2006

    • Author(s)
      Tsurimoto T
    • Total Pages
      24
    • Publisher
      Cold Spring Harbor Press NY
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] PCNA-interacting proteins(in Proliferating Cell Nuclear Antigen (PCNA) ed.Lee H.)2006

    • Author(s)
      Tsurimoto T.
    • Total Pages
      25
    • Publisher
      Research Signpost, Kerara, India
    • Related Report
      2006 Annual Research Report
  • [Book] PCNA and RFC proteins in Genomic Stability. (in DNA Replication in Eukaryotic Cells (2~<nd> Edition)ed. Depamphilis,M.L,)2006

    • Author(s)
      Tsurimoto T.
    • Total Pages
      24
    • Publisher
      Cold Spring Harbor Press NY
    • Related Report
      2006 Annual Research Report
  • [Book] PCNA and RFC proteins in Genomic Stability, (in DNA Replication in Eukaryotic Cells (2^<nd> Edition)(ed.DePamphilis, M.L.)2006

    • Author(s)
      Tsurimoto, T.
    • Publisher
      Cold Spring Harbor Press(New York)(In press)
    • Related Report
      2005 Annual Research Report
  • [Book] 複製フォーク複合体(細胞腫期イラストマップ 中山敬一編)2005

    • Author(s)
      釣本 敏樹
    • Total Pages
      7
    • Publisher
      羊土社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Remarks]

    • URL

      http://seibutsu.biology.kyushu-u.ac.jp/~chromosome/

    • Related Report
      2007 Annual Research Report

URL: 

Published: 2005-04-01   Modified: 2016-04-21  

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