Mechanistic insight into linkage between genome replication and particle formation of hepatitis C virus

• Project/Area Number: 18H02661
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49060:Virology-related
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: Suzuki Tetsuro 浜松医科大学, 医学部, 教授 (00250184)
• Co-Investigator(Kenkyū-buntansha): 鈴木 亮介 国立感染症研究所, ウイルス第二部, 室長 (50342902) ; 伊藤 昌彦 浜松医科大学, 医学部, 助教 (50385423)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000); Fiscal Year 2020: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2018: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: ウイルス複製 / タンパク質リン酸化 / 脱リン酸化酵素 / C型肝炎ウイルス / ゲノム複製 / 粒子形成 / ウイルス

Outline of Final Research Achievements

The molecular mechanisms of replication of plus-stranded RNA viruses, such as hepatitis C virus (HCV), have basically been analyzed by focusing on each process of the life cycle. In this study, we aimed to clarify the temporal and spatial transition for virus entry-genome replication-particle formation, and to identify the molecular mechanisms that contribute to the switching between the replication and particle formation.The following results were obtained: 1) Based on our previously established monitoring system for HCV replication, a real-time monitoring system for the processes of cell entry through genome replication was developed. 2) NS5A is likely to be a major viral factor involved in the control of the transition between genome replication and particle formation. 3) PPM1B variant 1 was identified as an enzyme that dephosphorylates NS5A. 4) LARP6, a member of the La family, was identified as a novel host factor that positively regulates the HCV life cycle.

Academic Significance and Societal Importance of the Research Achievements

HCVを含めプラス鎖RNAウイルスではゲノム複製と粒子形成がリンクする可能性が示されているがその調節機構は全く不明である。本研究はHCVをモデルとした解析であるが、近縁のペスチウイルスには同一機能が推定されるNS5Aが存在するのを初め、種々のウイルスの調節タンパク質がリン酸化によって機能制御されることが知られている。本研究が端緒となり、ウイルス増殖過程の制御において可逆的リン酸化がどのように関与するかの解析が多くのウイルスで進展することが期待される。本研究は、ウイルス学研究としてだけでなく、脱リン酸化酵素PPMファミリーの制御機構の解明にもつながる創造性を有する。

Research Products

• [Journal Article] BAC-DROP: Rapid Digestion of Proteome Fractionated via Dissolvable Polyacrylamide Gel Electrophoresis and Its Application to Bottom-Up Proteomics Workflow. (2021)
  • Author(s): Takemori A, Ishizaki J, Nakashima K, Shibata T, Kato H, Kodera Y, Suzuki T, Hasegawa H, Takemori N.
  • Journal Title: Journal of proteome research Volume: 20 Pages: 1535-1543
  • Peer Reviewed
• [Journal Article] Advancement of capture immunoassay for real-time monitoring of hepatitis E virus-infected monkey (2020)
  • Author(s): Khoris Indra Memdi、Chowdhury Ankan Dutta、Li Tian-Cheng、Suzuki Tetsuro、Park Enoch Y.
  • Journal Title: Analytica Chimica Acta Volume: 1110 Pages: 64-71
  • DOI: 10.1016/j.aca.2020.02.020
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Occludin‐binding single‐chain variable fragment and antigen‐binding fragment antibodies prevent hepatitis C virus infection (2020)
  • Author(s): Shimizu Yoshimi、Shinoda Takehiro、Shirasago Yoshitaka、Kondoh Masuo、Shinya Naoko、Hanada Kentaro、Yagi Kiyohito、Suzuki Tetsuro、Wakita Takaji、Kimura‐Someya Tomomi、Shirouzu Mikako、Fukasawa Masayoshi
  • Journal Title: FEBS Letters Volume: 595 Issue: 2 Pages: 220-229
  • DOI: 10.1002/1873-3468.13975
  • Peer Reviewed / Open Access
• [Journal Article] Controlling distance, size and concentration of nanoconjugates for optimized LSPR based biosensors. (2020)
  • Author(s): Chowdhury AD, Nasrin F, Gangopadhyay R, Ganganboina AB, Takemura K, Kozaki I, Honda H, Hara T, Abe F, Park S, Suzuki T, Park EY.
  • Journal Title: Biosens Bioelectron Volume: 170 Pages: 112657-112657
  • DOI: 10.1016/j.bios.2020.112657
  • NAID: 120006887333
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hollow magnetic-fluorescent nanoparticles for dual-modality virus detection. (2020)
  • Author(s): Ganganboina AB, Chowdhury AD, Khoris IM, Doong RA, Li TC, Hara T, Abe F, Suzuki T, Park EY.
  • Journal Title: Biosens Bioelectron Volume: 170 Pages: 112680-112680
  • DOI: 10.1016/j.bios.2020.112680
  • NAID: 120006889998
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] N‐terminal PreS1 Sequence Regulates Efficient Infection of Cell Culture‐generated Hepatitis B Virus (2020)
  • Author(s): Murayama A、Yamada N、Osaki 、Shiina 、Aly H H、Iwamoto M、Tsukuda S、Watashi K、Matsuda M、Suzuki R、Tanaka T、Moriishi K、Suzuki T、Nishitsuji H、Sugiyama M、Mizokami M、Shimotohno K、Wakita T、Muramatsu M、Liang T J、Kato T
  • Journal Title: Hepatology Volume: 「－」 Issue: 2 Pages: 520-532
  • DOI: 10.1002/hep.31308
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Prevalence and viral loads of polyomaviruses BKPyV, JCPyV, MCPyV, TSPyV and NJPyV and hepatitis viruses HBV, HCV and HEV in HIV-infected patients in China. (2020)
  • Author(s): Zhou X, Nakashima K, Ito M, Zhang X, Sakai S, Feng C, Sun H, Chen H, Li T-C, Suzuki T.
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 17066-17066
  • DOI: 10.1038/s41598-020-74244-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Agglutination of human polyomaviruses by using a tetravalent glycocluster as a cross-linker (2020)
  • Author(s): Ogata M, Onoda T, Koizumi A, Tokunaga Y, Ohta I, Nukuzuma S, Park EY, Usui T, Suzuki T
  • Journal Title: ACS Omega Volume: 5 Issue: 34 Pages: 21940-21947
  • DOI: 10.1021/acsomega.0c03269
  • Peer Reviewed / Open Access
• [Journal Article] Surfeit 4 contributes to the replication of hepatitis C virus using double membrane vesicles. (2020)
  • Author(s): Kong L, Aoyagi H, Yang Z, Ouyang T, Matsuda M, Fujimoto A, Watashi K, Suzuki R, Arita M, Yamagoe S, Dohmae N, Suzuki T, Suzuki T, Muramatsu M, Wakita T, Aizaki H.
  • Journal Title: Journal of Virology Volume: 94 Issue: 2
  • DOI: 10.1128/jvi.00858-19
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Characterization of the self-assembly of New Jersey polyomavirus VP1 into virus-like particles and the virus seroprevalence in Japan (2019)
  • Author(s): Zhou X, Bai H, Kataoka M, Ito M, Muramatsu M, Suzuki T, Li TC.
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 13085-13085
  • DOI: 10.1038/s41598-019-49541-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Activation of protein kinase R by hepatitis C virus RNA-dependent RNA polymerase. (2019)
  • Author(s): Suzuki R, Matsuda M, Shimoike T, Watashi K, Aizaki H, Kato T, Suzuki T, Muramatsu M, Wakita T.
  • Journal Title: Virology Volume: 529 Pages: 226-233
  • DOI: 10.1016/j.virol.2019.01.024
  • Peer Reviewed
• [Journal Article] USP15 Participates in Hepatitis C Virus Propagation through Regulation of Viral RNA Translation and Lipid Droplet Formation (2019)
  • Author(s): Kusakabe S、Suzuki T、Sugiyama Y、Haga S、Horike K、Tokunaga M、Hirano J、Zhang H、Chen DV、Ishiga H、Komoda Y、Ono C、Fukuhara T、Yamamoto M、Ikawa M、Satoh T、Akira S、Tanaka T、Moriishi Ki、Fukai M、Taketomi A、Yoshio S、Kanto T、Suzuki T、Okamoto T、Matsuura Y
  • Journal Title: Journal of Virology Volume: 93 Issue: 6 Pages: 1708-1718
  • DOI: 10.1128/jvi.01708-18
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The osteopontin-CD44 axis in hepatic cancer stem cells regulates IFN signaling and HCV replication. (2018)
  • Author(s): Shirasaki T, Honda M, Yamashita T, Nio K, Shimakami T, Shimizu R, Nakasyo S, Murai K, Shirasaki N, Okada H, Sakai Y, Sato T, Suzuki T, Yoshioka K, Kaneko S.
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 13143-13143
  • DOI: 10.1038/s41598-018-31421-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Monoclonal Antibodies against Occludin Completely Prevented Hepatitis C Virus Infection in a Mouse Model (2018)
  • Author(s): Shimizu Yoshimi、Shirasago Yoshitaka、Kondoh Masuo、Suzuki Tetsuro、Wakita Takaji、Hanada Kentaro、Yagi Kiyohito、Fukasawa Masayoshi
  • Journal Title: Journal of Virology Volume: 92 Issue: 8
  • DOI: 10.1128/jvi.02258-17
  • Peer Reviewed
• [Journal Article] Thermostable hepatitis C virus JFH1-derived variant isolated by adaptation to Huh7.5.1 cells (2018)
  • Author(s): Shirasago Yoshitaka、Fukazawa Hidesuke、Aizaki Hideki、Suzuki Tetsuro、Suzuki Takeru、Sugiyama Kazuo、Wakita Takaji、Hanada Kentaro、Abe Ryo、Fukasawa Masayoshi
  • Journal Title: Journal of General Virology Volume: 99 Issue: 10 Pages: 1407-1417
  • DOI: 10.1099/jgv.0.001117
  • Peer Reviewed
• [Journal Article] Molecular Basis of Encapsidation of Hepatitis C Virus Genome. (2018)
  • Author(s): Shi G, Suzuki T.
  • Journal Title: Front. Microbiol. Volume: 9 Pages: 396-396
  • DOI: 10.3389/fmicb.2018.00396
  • Peer Reviewed / Open Access / Int'l Joint Research