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Development of novel retroviral vectors showing no gene silencing effect

Research Project

Project/Area Number 22580104
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied biochemistry
Research InstitutionIshikawa Prefectural University (2012)
Nagoya University (2010-2011)

Principal Investigator

MIYAKE Katsuhide  石川県立大学, 生物資源環境学部, 教授 (90252254)

Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
KeywordsSUMO / インテグラーゼ / レトロウイルス / PIASy / Pc2 / MLV / PML / サイレンシング / HIV
Research Abstract

Pulldown assay and coimmunoprecipitation of cell extracts in which the integrase or reverse transcriptase of Moloney murine leukemia virus was transiently expressed showed that both enzymes interacted with PML proteins. In infected cells, interaction between the integrase and PML was also observed. Transient expression of PIASy and SUMO proteins facilitated SUMOylation of the integrase but had no apparent effects on the interaction with PML. A FLAG-tagged integrase colocalized with PML protein possibly in the PML body. Knockdown of PML by small interfering RNA resulted in reduced viral cDNA levels and integration efficiency. This suggested that PML proteins activated reverse transcription. Furthermore, we discovered direct interaction between the integrase and polycomb protein, Pc2.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (18 results)

All 2012 2011 2010

All Journal Article (5 results) (of which Peer Reviewed: 3 results) Presentation (11 results) Book (2 results)

  • [Journal Article] Moloney murine leukemia virus integrase and reverse transcriptase interact with PML proteins2012

    • Author(s)
      Y. Okino, Y. Inayoshi, Y. Kojima, S.Kidani, H. Kaneoka, A. Honkawa, K. Nishijima, K. Miyake, and S. Iijima
    • Journal Title

      J. Biochem

      Volume: 152 Pages: 161-169

    • NAID

      10031166125

    • Related Report
      2012 Final Research Report
  • [Journal Article] Chiken oviduct-specific expression of transgene by a hybrid ovalbumin enhancer and the Tet expression system2012

    • Author(s)
      D. Kodama, D. Nishimiya, K. Nishijima, Y. Okino, Y. Inayoshi, Y. Kojima, K. Ono, M. Motono, K. Miyake, Y. Kawabe, K. Kyogoku, T. Yamashita, M. Kamihira, and S. Iijima.
    • Journal Title

      J. Biosci. Bioeng

      Volume: 113 Pages: 146-153

    • Related Report
      2012 Final Research Report
  • [Journal Article]2012

    • Author(s)
      Akifumi Mizutani, Hiroyuki Tsunashima, Ken-ichi Nishijima, Takako Sasamoto Yuki Yamada, Yasuhiro Kojima, Makoto Motono, Jun Kojima, Yujin Inayoshi, Katsuhide Miyake, Enoch Y. Park, Shinji Iijima
    • Journal Title

      Transgenic Research

      Volume: vol.21 Issue: 1 Pages: 63-75

    • DOI

      10.1007/s11248-011-9511-0

    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Constitutive expression of the brgl gene requires GC-boxes neaz to the transcriptional start site2010

    • Author(s)
      Toshinari Itoh, Katsuhide Miyake, Takafumi Yamaguchi, Maasa Tsuge, Hidenori Kaneoka, Shinji Iijima
    • Journal Title

      The Journal of Biochemistry

      Volume: 149 Pages: 301-309

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Transcription Factor YY1 Interacts with Retroviral Integrases and Facilitates Integration of Moloney Murine Leukemia Virus cDNA into the Host Chromosomes2010

    • Author(s)
      Yujin Inayoshi, Yuuki Okino, Katsuhide Miyake, Akifumi Mizutani, Junko Yamamoto-Kishikawa, Yuya Kinoshita, Yusuke Morimoto, Kazuhito Imamura, Mahboob Morshed, KenKono, Toshinari Itoh, Ken-ichi Nishijima, Shinji Iijima
    • Journal Title

      Journal of Virology

      Volume: 84 Pages: 8250-8261

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] レトロウイルスインテグラーゼと核内タンパク質PMLの相互作用とその機能2012

    • Author(s)
      三宅克英、沖野雄気、小島佑介、木渓俊介、西島謙一、飯島信司
    • Organizer
      日本農芸化学会2012年度大会
    • Place of Presentation
      京都女子大学(京都府)
    • Year and Date
      2012-03-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] レトロウイルスインテグラーゼと核内タンパク質PMLの相互作用とその機能2012

    • Author(s)
      三宅克英・沖野雄気・小島佑介・木渓俊介・西島謙一・飯島信司
    • Organizer
      日本農芸化学会2012年度大会
    • Place of Presentation
      京都女子大学
    • Related Report
      2012 Final Research Report
  • [Presentation] The new target of sumoylation on UV irradiation2012

    • Author(s)
      M. Tsuge, H. Kaneoka, S. Kidani, Y. Masuda, K. Miyake and S.Iijima
    • Organizer
      The 25thAnnual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2012)
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2012 Final Research Report
  • [Presentation] UV 照射時における新たな SUMO 化の可能性2012

    • Author(s)
      柘植真亜沙・金岡英徳・木渓俊介・増田裕介・三宅克英・飯島信司
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場
    • Related Report
      2012 Final Research Report
  • [Presentation] The new target of sumoylation on UV irradiation2012

    • Author(s)
      Maasa Tsuge
    • Organizer
      The 25th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2012).
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2012 Annual Research Report
  • [Presentation] UV照射時における新たなSUMO化の可能性2012

    • Author(s)
      柘植真亜沙
    • Organizer
      第35回日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場
    • Related Report
      2012 Annual Research Report
  • [Presentation] レトロウイルスインテグラーゼのSUMO化とその役割2011

    • Author(s)
      沖野雄気、木渓俊介、小島佑介、三宅克英、稲吉勇仁、西島謙一、飯島信司
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学(東京都)
    • Year and Date
      2011-09-26
    • Related Report
      2011 Annual Research Report
  • [Presentation] レトロウイルスインテグラーゼのSUMO化とその役割2011

    • Author(s)
      沖野雄気、木溪俊介、小島佑介、三宅克英、稲吉勇仁、西島謙一、飯島信司
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学
    • Related Report
      2012 Final Research Report
  • [Presentation] ニワトリ卵白蛋白質におけるN結合型糖鎖付加能の解析2011

    • Author(s)
      西島謙一、水谷昭文、元野誠、綱島裕之、山田裕貴、三宅克英、飯島信司
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学
    • Related Report
      2012 Final Research Report
  • [Presentation] レトロウイルスゲノムのインテグレーションメカニズムの解析2010

    • Author(s)
      沖野雄気、稲吉勇仁、水谷昭文、三宅克英、西島謙一、飯島信司
    • Organizer
      第33回日本分子生物学会年会 第83回日本生化学会大会合同大会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2010-12-10
    • Related Report
      2010 Annual Research Report
  • [Presentation] レトロウイルスのゲノムインテグレーションメカニズムの解析2010

    • Author(s)
      三宅克英、沖野雄気、稲吉勇仁、水谷昭文、西島謙一、飯島信司
    • Organizer
      第62回日本生物工学会大会
    • Place of Presentation
      ワールドコンベンションセンターサミットフェニックス・シーガイア・リゾート(宮崎市山崎町浜山)
    • Year and Date
      2010-10-28
    • Related Report
      2010 Annual Research Report
  • [Book] トランスジェニックニワトリ作製のための新規DNA導入法の開発2012

    • Author(s)
      三宅克英 ・金岡英徳・西島謙一・飯島信司
    • Publisher
      agoya University e ブックシリーズ
    • Related Report
      2012 Final Research Report
  • [Book] トランスジェニックニワトリ作製のための新規DNA導入法の開発2012

    • Author(s)
      三宅克英
    • Publisher
      Nagoya University eブックシリーズ
    • Related Report
      2012 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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