Development of novel retroviral vectors showing no gene silencing effect

• Project/Area Number: 22580104
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Ishikawa 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: ¥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)
• Keywords: SUMO / 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.

Research Products

• [Journal Article] Moloney murine leukemia virus integrase and reverse transcriptase interact with PML proteins (2012)
  • 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
• [Journal Article] Chiken oviduct-specific expression of transgene by a hybrid ovalbumin enhancer and the Tet expression system (2012)
  • 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
• [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
  • Peer Reviewed
• [Journal Article] Constitutive expression of the brgl gene requires GC-boxes neaz to the transcriptional start site (2010)
  • 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
  • Peer Reviewed
• [Journal Article] Transcription Factor YY1 Interacts with Retroviral Integrases and Facilitates Integration of Moloney Murine Leukemia Virus cDNA into the Host Chromosomes (2010)
  • 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
  • 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
• [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
• [Presentation] The new target of sumoylation on UV irradiation (2012)
  • 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
• [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
• [Presentation] The new target of sumoylation on UV irradiation (2012)
  • 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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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