A comprehensive protein interaction analysis for developing new therapeutic targets in HIV infection using cell-free protein synthesis technology

• Project/Area Number: 24390115
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Virology
• Research Institution: Yokohama City University
• Principal Investigator: RYO Akihide 横浜市立大学, 医学(系)研究科(研究院), 教授 (20363814)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2014: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2013: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: ウイルス / タンパク質 / プロテオーム / 酵素 / バイオテクノロジー / HIV / 宿主因子 / 蛋白質

Outline of Final Research Achievements

In this study, we took advantage of the comprehensive protein interaction analysis using cell-free protein synthesis technology. We established an assay system for monitoring the functional interaction between HIV accessory proteins and intrinsic anti-virus host factors. Using this system, we found that apoptosis signal-regulating kinase 1 (ASK1) interferes with the counteraction of Vif and revitalizes the A3G-mediated viral restriction. ASK1 binds the BC-box of Vif, thereby disrupting the assembly of the Vif-ubiquitin ligase complex. Consequently, ASK1 stabilizes A3G and promotes its incorporation into viral particles, ultimately reducing viral infectivity. Furthermore, treatment with the antiretroviral AZT (zidovudine) induces ASK1 expression and restores the antiviral activity of A3G in HIV-1-infected cells. This study thus demonstrates a distinct function of ASK1 in restoring the host antiviral system that can be enhanced by AZT treatment.

Research Products

• [Journal Article] ASK1 restores the antiviral activity of APOBEC3G by disrupting HIV-1 Vif-mediated counteraction. (2015)
  • Author(s): Miyakawa K, Matsunaga S, Kanou K, Matsuzawa A, Morishita R, Kudoh A, Shindo K, Yokoyama M, Sato H, Kimura H, Tamura T, Yamamoto N, Ichijo H, Takaori-Kondo A, Ryo A.
  • Journal Title: Nature Communications Volume: 6:6945 Issue: 1 Pages: 6945-6945
  • DOI: 10.1038/ncomms7945
  • NAID: 120005745628
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Quantitative analysis of location- and sequence-dependent deamination by APOBEC3G using real-time NMR spectroscopy (2014)
  • Author(s): Furukawa, A., Sugase, K., Morishita, R., Nagata, T., Kodaki, T., Takaori-Kondo, A., Ryo, A., Katahira, M.
  • Journal Title: Angew. Chem. Int. Ed. Engl. Volume: 53 Issue: 9 Pages: 2349-2352
  • DOI: 10.1002/anie.201309940
  • NAID: 120005647511
  • Peer Reviewed / Open Access
• [Journal Article] The phosphorylation of HIV-1 Gag by atypical protein kinase C facilitates viral infectivity by promoting Vpr incorporation into virions. (2014)
  • Author(s): Kudoh A, Takahama S, Sawasaki T, Ode H, Yokoyama M, Okayama A, Ishikawa A, Miyakawa K, Matsunaga S, Kimura H, Sugiura W, Sato H, Hirano H, Ohno S, Yamamoto N, Ryo A
  • Journal Title: Retrovirology Volume: 11 Issue: 1 Pages: 9-9
  • DOI: 10.1186/1742-4690-11-9
  • Peer Reviewed / Open Access
• [Journal Article] Generation of rhesus macaque-tropic HIV-1 clones that are resistant to major anti-HIV-1 restriction factors (2013)
  • Author(s): Nomaguchi M, Yokoyama M, Kono K, Nakayama EE, Shioda T, Doi N, Fujiwara S, Saito A, Akari H, Miyakawa K, Ryo A, Ode H, Iwatani Y, Miura T, Igarashi T, Sato H, Adachi A
  • Journal Title: J Virol Volume: 87(21) Issue: 21 Pages: 11447-11461
  • DOI: 10.1128/jvi.01549-13
  • Peer Reviewed
• [Journal Article] Interferon-induced SCYL2 limits release of HIV-1 by triggering PP2A-mediated dephosphorylation of the viral protein Vpu. (2012)
  • Author(s): Miyakawa K., et al.,
  • Journal Title: Sci Signal. Volume: 5 Issue: 245
  • DOI: 10.1126/scisignal.2003212
  • Peer Reviewed
• [Journal Article] NMR study of xenotropic murine leukemia virus-related virus protease in a complex with amprenavir (2012)
  • Author(s): Ayako Furukawa, Hideyasu Okamura, Ryo Morishita, Satoko Matsunaga, Naohiro Kobayashi, Takahisa Ikegami, Tsutomu Kodaki, Akifumi Takaori-Kondo, Ryo Akihide, Takashi Nagata, Masato Katahira
  • Journal Title: Biochem Biophys Res Commun. Volume: 24巻 Issue: 2 Pages: 284-289
  • DOI: 10.1016/j.bbrc.2012.07.083
  • NAID: 120004873605
  • Peer Reviewed
• [Journal Article] Molecular and enzymatic characterization of XMRV protease by a cell-free proteolytic analysis. (2012)
  • Author(s): Matsunaga S
  • Journal Title: J Proteomics Volume: 75 Issue: 15 Pages: 4863-4873
  • DOI: 10.1016/j.jprot.2012.05.047
  • Peer Reviewed
• [Presentation] Newly developed measles viral vector can efficiently transduce multiple genes into naïve T cells. (2014)
  • Author(s): Takafumi Hiramoto et al.
  • Organizer: 56th American Society of Hematology Annual Meeting & Exposition
  • Place of Presentation: Moscone Center(San Francisco)
  • Year and Date: 2014-12-06 – 2014-12-09
• [Presentation] 宿主防御因子Tetherin/BST2によるB型肝炎ウイルス感染抑制とその回避機構の解明. (2014)
  • Author(s): 宮川 敬, 松永智子, 渡士幸一, 杉山真也, 溝上雅史, 脇田隆字,梁 明秀
  • Organizer: 第62回日本ウイルス学会学術集会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2014-11-10 – 2014-11-12
• [Presentation] H11/HSPB8 confers HIV resistance to Human placental trophoblasts. (2014)
  • Author(s): Ayumi Kudoh, Kei Miyakawa, Satoko Matsunaga, Isao Kosugi, Ryo Akihide.
  • Organizer: The 13th Awaji International Forum on Infection and Immunity in Nara
  • Place of Presentation: 奈良県新公会堂（奈良県）
  • Year and Date: 2014-09-23 – 2014-09-26
• [Presentation] Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type3 for generation and characterization of monoclonal antibody. (2014)
  • Author(s): Satoko Matsunaga, Ayumi Kudoh, Hirokazu Kimura, Akihide Ryo.
  • Organizer: Protein Island Matsuyama (PIM) International Symposium 2014
  • Place of Presentation: 愛媛大学（愛媛県）
  • Year and Date: 2014-09-15 – 2014-09-17
• [Presentation] HIV-1 ViI-AP0BEC3Gの相互作用を調節する新規宿主因子の同定 (2012)
  • Author(s): 宮川 敬
  • Organizer: 第26回日本エイズ学会学術集会・総会
  • Place of Presentation: 慶應義塾大学(神奈川)
  • Year and Date: 2012-11-26
• [Presentation] HIVアクセサリー蛋白質Vpxと相互作用する宿主因子の網羅的探索と機能解析 (2012)
  • Author(s): 工藤 あゆみ
  • Organizer: 第26回日本エイズ学会学術集会・総会
  • Place of Presentation: 慶應義塾大学(神奈川)
  • Year and Date: 2012-11-24
• [Presentation] HIV-1 Vpuの翻訳後修飾とその意義 (2012)
  • Author(s): 佐藤 遥
  • Organizer: 第26回日本エイズ学会学術集会・総会
  • Place of Presentation: 慶應義塾大学(神奈川)
  • Year and Date: 2012-11-24
• [Presentation] typical protein kinase CによるHIV-1 Gag Ser487のリン酸化はウイルス感染性を増強する (2012)
  • Author(s): 工藤 あゆみ
  • Organizer: 第60回日本ウイルス学会学術集会
  • Place of Presentation: グランキューブ大阪(大阪)
  • Year and Date: 2012-11-13
• [Presentation] Vpxのリン酸化はHIV感染を制御する．
  • Author(s): 宮川 敬、松永智子、工藤あゆみ、梁 明秀
  • Organizer: 第27回日本エイズ学会学術集会・総会
  • Place of Presentation: 熊本市民会館（熊本）
• [Presentation] Vpxの機能抑制的に作用する宿主因子の同定とその作用機序．
  • Author(s): 工藤あゆみ、宮川 敬、松永智子、小杉伊三夫、梁 明秀
  • Organizer: 第61回日本ウイルス学会学術集会
  • Place of Presentation: 神戸国際会議場（兵庫）
• [Presentation] Wheat germ cell-free protein production of human immunodeficiency virus accessory protein Vif.
  • Author(s): Mayu Miyamoto, Satoko Matsunaga, Kei Miyakawa, Akihide Ryo
  • Organizer: 第12回ヒトプロテオーム機構国際会議
  • Place of Presentation: パシフィコ横浜（神奈川）
• [Presentation] A proteomic approach to decipher the molecular link between HIV-1 Gag and host proteins．
  • Author(s): Akihide Ryo
  • Organizer: 第12回ヒトプロテオーム機構国際会議
  • Place of Presentation: パシフィコ横浜（神奈川）
• [Presentation] Identification and characterization of Vpx-interacting host protein kinases.
  • Author(s): Kei Miyakawa, Satoko Matsunaga, Ayumi Kudoh, Akihide Ryo
  • Organizer: CSHL Meeting, Retroviruses
  • Place of Presentation: Cold Spring Harbor Laboratory(America, New York)
• [Remarks] 横浜市立大学大学院医学研究科 微生物学・分子生体防御学
  • URL: https://www-user.yokohama-cu.ac.jp/~saikin/