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Studies translesion DNA synthesis complex

Research Project

Project/Area Number 16370077
Research Category

Grant-in-Aid for Scientific Research (B)

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

Principal Investigator

OHMORI Haruo  Kyoto University, Institute for Virus Research, Associate Professor, ウイルス研究所, 助教授 (10127061)

Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥15,400,000 (Direct Cost: ¥15,400,000)
Fiscal Year 2006: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2005: ¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 2004: ¥6,000,000 (Direct Cost: ¥6,000,000)
KeywordsTranslesion DNA synthesis / Mutation / DNA polymerases / Y-family / Pol kappa / Protein-protein interactions / Binding motif sequence / Protein secondary structures / DNA損傷 / 損傷バイパス複合体 / REV1 / proline / ペプチド / Polη / Polι / Polκ / PCNA / 複製の忠実度 / 校正機能
Research Abstract

We and other groups have shown that the three Y-family DNA polymerases Polη, Polι, and Polκ interact with a C-terminal region of REV 1, another member of the Y-family DNA polymerases. The same C-terminal region of hREV1 was also shown to interact with hREV7, a non-catalytic subunit of Polζ, which is another TLS DNA polymerase belonging to the B-family. This has suggested us that hREV1 plays a crucial role in cellular TLS events. Unlike for the PCNA binding, Polη, Polι, and Polκ did not seem to have an apparent consensus sequence for the hREV1-interaction. However, our further experiments revealed that short sequences less than 10 residues are sufficient for the hREV1-interaction in all of the three Pols. In addition, in all cases, two consecutive phenylalanine (FF) residues were found to be critical for the hREV1-interactions. No conserved amino acid residue is required for the N-terminal side of the FF motif, but the presence of several residues at its C-terminal side is essential while the sequence is not conserved. Using surface plasmon resonance method, we showed that short synthetic peptides of the sequence derived from the hREV1-binding sites of Polκ directly bind the C-terminal region of hREV1. Furthermore, such peptides did inhibit the binding between purified hREV1 and Polκ proteins.
To examine the biological relevance of the interaction between hREV1 and Polκ, we employed complementation assay using Polκ-deficient MEF (mouse embryonic fibroblast) cells, which exhibit increased sensitivity to BPDE (benzo[a[pyrene dihydroxydiol epoxide, the ultimate carcinogen of benzo[a[pyrene] and UV-irradiation. We observed that the sensitivities to BPDE or UV were recovered by transient expression of the wild-type of Polκ (in a fusion with GFP), but not by that of the FF567-578AA mutant deficient for the hREV1-binding.

Report

(4 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (33 results)

All 2007 2006 2005 2004 Other

All Journal Article (33 results)

  • [Journal Article] Translesion DNA synthesis across mono ADP-ribosylated dG by Y-family DNA polymerases.2007

    • Author(s)
      M.Kawanishi et al.
    • Journal Title

      New Developments in Mutation Research (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Integrating checkpoint signaling with transOlesion synthesis of bulky DNA adducts.2007

    • Author(s)
      L.Barkley, H.Ohmori, C.Vaziri
    • Journal Title

      Cell Biochemistry and Biophysics (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Translesion DNA synthesis across mono ADP-ribosylated dG by Y- family DNA polymerases.2007

    • Author(s)
      Kawanishi M, Matsukawa K, Ohashi E, Takamura T, Otsuka Y, Watanabe M, Sugimura T, Wakabayashi K, Hanaoka F, Ohmori H, Yagi T.
    • Journal Title

      New Developments in Mutation Research (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Integrating S-phase Checkpoint Signaling with Trans-Lesion Synthesis of Bulky DNA Adducts.2007

    • Author(s)
      Barkley LR, Ohmori H, Vaziri C.
    • Journal Title

      Cell Biochemistry and Biophysics (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Integrating S-phase checkpoint signaling with trans-lesion synthesis of bulky DNA addcuts2007

    • Author(s)
      Barkley LM, Ohmori H, Vaziri C.
    • Journal Title

      Cell Biochemistry and Biophysics (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Translesion DNA synthesis across mono ADP-ribosylated dG by Y-family DNA polymerases2007

    • Author(s)
      Kawanishi M, Matsukawa K, Ohashi E., Takamura T, 他7名
    • Journal Title

      New Developments in Mutation Research (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Rad 18 regulates DNA polymerase κ and is required for recovery from S-phase checkpoint-mediated arrest.2006

    • Author(s)
      X.Bi et al.
    • Journal Title

      Molecular and Cellular Biology 26

      Pages: 3527-3540

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Rad18 regulates DNA polymerase K and is required for recovery from S- phase checkpoint-mediated arrest.2006

    • Author(s)
      Bi X, Barkley LR, Slater DM, Tateishi S, Yamaizumi M, Ohmori H, Vaziri C.
    • Journal Title

      Mol. Cell. Biol. 26

      Pages: 3527-3540

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Rad18 regulates DNA polymerase k and is required for recovery from S-phase checkpoint-mediated arrest.2006

    • Author(s)
      Bi X, Barkley LR, Slater DM, Tateishi S, Yamaizumi M, Ohmori
    • Journal Title

      Molecular and Cellular Biology 26

      Pages: 3257-3540

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Rad18 regulates DNA polymerase kappa and is required for recovery from S-phase checkpoint mediated arrest2006

    • Author(s)
      X.Bi et al.
    • Journal Title

      Molecular and Cellular Biology 26(in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Translesion DNA synthesis across mono ADP-ribosylated dG by Y-family DNA polymerases.2006

    • Author(s)
      N.Kawanishi et al.
    • Journal Title

      Developments in Mutation Research (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Error-prone and inefficient replication across 8-hydroxyguanine (8-oxoguanine) in human and mouse ras gene fragments by DNA polymerase κ.2005

    • Author(s)
      P.Jaloszynski et al.
    • Journal Title

      Genes to Cells 10

      Pages: 543-550

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Normal immunoglobulin gene somatic hypermutation in Polk-double-deficient mice.2005

    • Author(s)
      T.Shimizu et al.
    • Journal Title

      Immunology Letters 9

      Pages: 259-264

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] DNA polymerase κ is specifically required for recovery from the benzo[α]pyrene-dihydrodiol epoxide (BPDE)-induced S-phase checkpoint2005

    • Author(s)
      X.Bi et al.
    • Journal Title

      Journal of Biological Chemistry 280

      Pages: 22343-22355

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Error-prone and inefficient replication across 8- hydroxyguanine (8-oxoguanine) in human and mouse ras gene fragments by DNA polymerase κ.2005

    • Author(s)
      Jaloszynski P, Ohashi E, Ohmori H, Nishimura S.
    • Journal Title

      Genes Cells. 10

      Pages: 543-550

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Normal immunoglobulin gene somatic hypermutation in Polk-Poli double-deficient mice.2005

    • Author(s)
      Shimizu T, Azuma T, Ishiguro M, Kanjo N, Yamada S. Ohmori H.
    • Journal Title

      Immunol Lett. 9

      Pages: 259-264

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Normal immunoglobulin gene somatic hypermutation in Polk-Poli double deficient mice.2005

    • Author(s)
      T.Shimizu et al.
    • Journal Title

      Immunological Letters 9

      Pages: 259-264

    • Related Report
      2005 Annual Research Report
  • [Journal Article] DNA polymerase kappa is specifically required for recovery from the benzo[a]pyrene-dihydrodiol expoxide2005

    • Author(s)
      X.Bi et al.
    • Journal Title

      J.Biological Chemistry 280

      Pages: 22334-22355

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Error-prone and inefficient replication across 8-hydroxyguanine (8-oxoguanine) in human and mouse ras2005

    • Author(s)
      P.Jaloszynski et al.
    • Journal Title

      Genes to Cells 10

      Pages: 543-550

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Normal immunoglobulin gene somatic hypermutation in Polκ-Polι double-deficient mice2005

    • Author(s)
      Shimizu, T., et al.
    • Journal Title

      Immunology Letters (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Translesion DNA synthesis across monoADP-ribosylated dG by Y-family DNA polymerases2005

    • Author(s)
      Kawanishi, M., et al.
    • Journal Title

      Progress in Mutation Research (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Error-prone and inefficient replication across 8-hydroxyguanine in human and mouse ras gene fragments by DNA polymerase κ.2005

    • Author(s)
      Jaloszynski, P., et al.
    • Journal Title

      Genes to Cells (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Mammalian Polκ : regulation of its expression and lesion substrates.2004

    • Author(s)
      H.Ohmori et al.
    • Journal Title

      Advance in Protein Chemictry 69

      Pages: 265-278

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Interaction of hREV1 with three human Y-family DNA polymerases.2004

    • Author(s)
      E.Ohashi et al.
    • Journal Title

      Genes to Cells 9

      Pages: 523-531

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Translesion synthesis past equine estrogen-derived 2'-deoxycytidine DNA adducts by human DNA polymerases η and κ.2004

    • Author(s)
      N.Suzuki et al.
    • Journal Title

      Biochemistry 43

      Pages: 11312-11320

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Mutagenic properties of 3-(deoxyguanosine-N2-yl)-2-acetylaminofluorene, a persistent acetylaminofluorene-derived DNA adduct in mammalian cells.2004

    • Author(s)
      M.Yasui et al.
    • Journal Title

      Biochemistry 43

      Pages: 15005-15013

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Mammalian Polκ : regulation of its expression and lesion substrates.2004

    • Author(s)
      Ohmori H, Ohashi E, Ogi T.
    • Journal Title

      Adv.Protein Chem 69

      Pages: 265-278

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Interaction of hREV1 with three human Y-family DNA polymerase.2004

    • Author(s)
      Ohashi E, Murakumo Y, Kanjo N, Akagi J, Masutani C, Hanaoka F, Ohmori H.
    • Journal Title

      Genes to Cells 9

      Pages: 523-531

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Translesion synthesis past equine estrogen-derived 2'-deoxycytidine DNA adducts by human DNA polymerases η and κ.2004

    • Author(s)
      Suzuki N, Yasui M, Santosh Laxmi YR, Ohmori H, Hanaoka F, Shibutani S.
    • Journal Title

      Biochemistry 43

      Pages: 11312-11320

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Mutagenic properties of 3-(deoxyguanosin-N^2-yl)-2- acetylaminofluorene, a persistent acetylaminofluorene-derived DNA adduct in mammalian cells.2004

    • Author(s)
      Yasui M, Dong H, Bonala RR, Suzuki N, Ohmori H, Hanaoka F, Johnson F, Grollman AP, Shibutani S.
    • Journal Title

      Biochemistry 43

      Pages: 15005-15013

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Interaction of hREV1 with three human Y-family DNA polymerases.2004

    • Author(s)
      Ohashi, E., et al.
    • Journal Title

      Genes to Cells 9

      Pages: 523-531

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Mammalian Polκ : regulation of its expression and lesion substrates.2004

    • Author(s)
      Ohmori, H., et al.
    • Journal Title

      Advances in Protein Chemistry 69

      Pages: 265-278

    • Related Report
      2004 Annual Research Report
  • [Journal Article] DNA Polymerase K is specifically required for recovery from the benzo[a]pyrene-dihydrodiol epoxide (BPDE)-induced S-phase checkpoint.

    • Author(s)
      Bi X, Slater DM, Ohmori H, Vaziri C.
    • Journal Title

      J.Biol.Chem. 280

      Pages: 22343-22355

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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