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Effects of DNA-protein cross-link lesions on eukaryotic transcription and replication

Research Project

Project/Area Number 23710070
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Risk sciences of radiation/Chemicals
Research InstitutionHiroshima University

Principal Investigator

NAKANO Toshiaki  広島大学, 大学院・理学研究科, 助教 (10526122)

Project Period (FY) 2011 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
KeywordsDNA損傷 / 転写 / 複製
Research Abstract

DNA-protein cross-links (DPCs) are superbulky and ubiquitous DNA lesions induced by various agents. It is likely that DPCs interfere with many aspects of DNA transactions. In the present study, we established in vitroassays to investigate the effect ofDPCs on eukaryotic transcription catalyzed by RNA polymerase II, showing that the formation of runoff products are dependent on the amount of nuclear cell extracts and reaction time. We then analyzed the effect of a model bulky lesion (fluorescein) on transcription. Fluorescein on the transcribed strand but not on the nontrascribed strand inhibited the transcription. We also established a novel method to introduce DPCs by employing Click reactions between protein azides and C5 alkyne-dT. We are currently studying the effect of DPCs on transcription by RNA polymerase II.

Report

(3 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Research-status Report
  • Research Products

    (14 results)

All 2013 2012 2011 Other

All Journal Article (5 results) (of which Peer Reviewed: 5 results) Presentation (8 results) Remarks (1 results)

  • [Journal Article] Translocation and stability of replicative DNA helicase upon encountering DNA-protein cross-links.2013

    • Author(s)
      Nakano, T., Miyamoto-Matsubara M., Shoulkamy M.I. Amir M.H., Pack S.P., Ishimi Y., and Ide, H.
    • Journal Title

      J. Biol. Chem.

      Volume: 288 Pages: 4649-4658

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] T7 RNA polymerases backed up by covalently trapped proteins catalyze highly error prone transcription.2012

    • Author(s)
      Nakano, T., Ouchi R., Kawazoe, J., Pack S.P., Makino K., and Ide, H.
    • Journal Title

      J. Biol. Chem.

      Volume: 287 Pages: 6562-6572

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Detection of DNA-protein crosslinks (DPCs) by novel direct fluorescence labeling methods: distinct stabilities of aldehyde and radiation-induced DPCs2012

    • Author(s)
      Shoulkamy M.I., Nakano T., Ohshima M., Hirayama R., Uzawa A., Furusawa Y., and Ide H.
    • Journal Title

      Nucleic Acids Res

      Volume: 40

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] T7 RNApolymerases backed up by covalently trapped proteins catalyze highly errorprone transcription2012

    • Author(s)
      Nakano T., Ouchi R., Kawazoe J., PackS. P., Makino K., and Ide H.
    • Journal Title

      J. Biol. Chem.

      Volume: 287 Issue: 9 Pages: 6562-6572

    • DOI

      10.1074/jbc.m111.318410

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] T7 RNA polymerases backed up by covalently trapped proteins catalyze highly error prone transcription.2012

    • Author(s)
      Toshiaki Nakano, Ryo Ouchi, Junya Kawazoe, Seung Pil Pack, Keisuke Makino and Hiroshi Ide
    • Journal Title

      The Journal of Biological Chemistry

      Volume: 287(9) Pages: 6562-6572

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Presentation] DNA-タンパク質クロスリンク損傷が誘発する非標的転写エラー2012

    • Author(s)
      中野敏彰, (他3名)
    • Organizer
      第33回日本環境変異原学会
    • Place of Presentation
      静岡市
    • Related Report
      2012 Final Research Report
  • [Presentation] DNA-protein crosslinks stall T7 RNA polymerase and induce untargeted transcriptional mutations2012

    • Author(s)
      Toshiaki Nakano, (他5名)
    • Organizer
      12th International Workshop on Radiation Damage to DNA
    • Place of Presentation
      Prague, Czech
    • Related Report
      2012 Final Research Report
  • [Presentation] DNA-protein crosslinks stall T7 RNA polymerases and induce untargeted transcriptional mutations2012

    • Author(s)
      Nakano T., Ouchi R.,Miyamoto-Matsubara M., Shoulkamy M.I., Amir M.H., and Ide H.
    • Organizer
      12th International Workshop on Radiation Damage to DNA
    • Place of Presentation
      チェコ(プラハ)
    • Related Report
      2012 Annual Research Report
  • [Presentation] DNA-タンパク質クロスリンク損傷が誘発する非標的転写エラー2012

    • Author(s)
      中野敏彰,大内 綾,宮本(松原)真由美,井出 博
    • Organizer
      日本環境変異原学会
    • Place of Presentation
      静岡県
    • Related Report
      2012 Annual Research Report
  • [Presentation] 蛍光標識を用いたDNA-タンパク質クロスリンク損傷が複製ヘリカーゼの進行に及ぼす影響2011

    • Author(s)
      中野敏彰, (他5名)
    • Organizer
      第34回日本分子生物学会
    • Place of Presentation
      横浜市
    • Year and Date
      2011-12-15
    • Related Report
      2012 Final Research Report
  • [Presentation] 蛍光標識を用いたDNA-タンパク質クロスリンク損傷の新規検出法2011

    • Author(s)
      中野敏彰, (他4名)
    • Organizer
      第32回日本環境変異原学会
    • Place of Presentation
      東京
    • Year and Date
      2011-11-21
    • Related Report
      2012 Final Research Report
  • [Presentation] 蛍光標識を用いたDNA-タンパク質クロスリンク損傷の新規検出法2011

    • Author(s)
      中野 敏彰、Mahmoud Shoulkamy、大嶌 麻妃子、宮本(松原) 真由美、井出  博
    • Organizer
      日本環境変異原学会
    • Place of Presentation
      東京
    • Related Report
      2011 Research-status Report
  • [Presentation] DNA-タンパク質クロスリンク損傷が複製ヘリカーゼの進行に及ぼす影響2011

    • Author(s)
      中野敏彰, 宮本(松原) 真由美, 澤和 美菜子, 石見 幸男, 井出博
    • Organizer
      日本分子生物学会
    • Place of Presentation
      神奈川県(横浜)
    • Related Report
      2011 Research-status Report
  • [Remarks] 遺伝子化学研究室ホームページ

    • URL

      http://home.hiroshima-u.ac.jp/genechem/index.html

    • Related Report
      2012 Final Research Report

URL: 

Published: 2011-08-05   Modified: 2019-07-29  

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