Analysis of the replication mechanism of hepatitis C virus in membrane vesicles containing replication complexes

2018 Fiscal Year Annual Research Report

• Project/Area Number: 15K09034
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: 相崎 英樹 国立感染症研究所, ウイルス第二部, 室長 (00333360)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Keywords: HCV / 複製 / 膜小胞 / プラス鎖RNAウイルス / 複製複合体

Outline of Annual Research Achievements

C型肝炎ウイルス(HCV)、ポリオウイルス、サーズコロナウイルス等のプラス鎖RNAウイルスは複製複合体を含む膜小胞(Double membrane vesicle(DMV))を形成し、その中で複製複合体を形成し、複製することが知られている。本研究ではHCVの複製に必須な膜小胞形成メカニズムの解析を行った。
HCVNS4BはHCV複製に関与する膜小胞の形成に重要な役割を果たしていることが知られているので、NS4Bと結合する膜タンパクを同定し、そのタンパクの膜小胞の形成における役割について解析した。プロテオーム解析およびsiRNAスクリーニングにて、NS4B結合膜タンパクとしてSurfeit 4 (Surf4)を見出した。SURF4はCOPIを介した小胞輸送（メンブレントラフィック）に関与するタンパクとして知られている。各種解析から、Surf4とNS4Bとの相互作用が複製に重要であることを示した。電顕観察でSurf4 siRNA処理細胞ではDMVの数が大きく減少していたことから、Surf4はDMV形成を介してHCV複製に関与している可能性が見出された。また、ポリオウイルスレプリコンをsiRNAで処理すると複製が減少し、デングウイルスでは減少しなかった。ポリオウイルスはDMVを形成し複製することが知られている、一方、デングウイルスではDMVを形成しないことから、Surf4はHCV、ポリオウイルス、サーズコロナウイルス等のプラス鎖RNAウイルスに共通の複製コンポーネント形成に作用する可能性が示された。
以上の結果から、Surf4がNS4Bと相互作用することによってDMVを形成し、HCV複製に関与していることを示した。

Research Products

• [Journal Article] Epidermal growth factor receptor is a host-entry cofactor triggering hepatitis B virus internalization (2019)
  • Author(s): Iwamoto Masashi、Saso Wakana、Sugiyama Ryuichi、Ishii Koji、Ohki Mio、Nagamori Shushi、Suzuki Ryosuke、Aizaki Hideki、Ryo Akihide、Yun Ji-Hye、Park Sam-Yong、Ohtani Naoko、Muramatsu Masamichi、Iwami Shingo、Tanaka Yasuhito、Sureau Camille、Wakita Takaji、Watashi Koichi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Pages: 8487～8492
  • DOI: 10.1073/pnas.1811064116
  • Peer Reviewed
• [Journal Article] Activation of protein kinase R by hepatitis C virus RNA-dependent RNA polymerase (2019)
  • Author(s): Suzuki Ryosuke、Matsuda Mami、Shimoike Takashi、Watashi Koichi、Aizaki Hideki、Kato Takanobu、Suzuki Tetsuro、Muramatsu Masamichi、Wakita Takaji
  • Journal Title: Virology Volume: 529 Pages: 226～233
  • DOI: 10.1016/j.virol.2019.01.024
  • Peer Reviewed
• [Journal Article] Regulation of Hepatitis C Virus Infection by Cellular Retinoic Acid Binding Proteins through the Modulation of Lipid Droplet Abundance (2019)
  • Author(s): Bang Bo-Ram、Li Meng、Tsai Kuen-Nan、Aoyagi Haruyo、Lee Shin-Ae、Machida Keigo、Aizaki Hideki、Jung Jae U.、Ou Jing-Hsiung James、Saito Takeshi
  • Journal Title: Journal of Virology Volume: 93 Pages: －
  • DOI: 10.1128/JVI.02302-18
  • Peer Reviewed
• [Journal Article] Troglitazone Impedes the Oligomerization of Sodium Taurocholate Cotransporting Polypeptide and Entry of Hepatitis B Virus Into Hepatocytes (2019)
  • Author(s): Fukano Kento、Tsukuda Senko、Oshima Mizuki、Suzuki Ryosuke、Aizaki Hideki、Ohki Mio、Park Sam-Yong、Muramatsu Masamichi、Wakita Takaji、Sureau Camille、Ogasawara Yuki、Watashi Koichi
  • Journal Title: Frontiers in Microbiology Volume: 9 Pages: －
  • DOI: 10.3389/fmicb.2018.03257
  • 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 Pages: 1407～1417
  • DOI: 10.1099/jgv.0.001117
  • Peer Reviewed
• [Journal Article] A Single Adaptive Mutation in Sodium Taurocholate Cotransporting Polypeptide Induced by Hepadnaviruses Determines Virus Species Specificity (2018)
  • Author(s): Takeuchi Junko S.、Fukano Kento、Iwamoto Masashi、Tsukuda Senko、Suzuki Ryosuke、Aizaki Hideki、Muramatsu Masamichi、Wakita Takaji、Sureau Camille、Watashi Koichi
  • Journal Title: Journal of Virology Volume: 93 Pages: －
  • DOI: 10.1128/JVI.01432-18
• [Journal Article] High-throughput neutralization assay for multiple flaviviruses based on single-round infectious particles using dengue virus type 1 reporter replicon (2018)
  • Author(s): Matsuda Mami、Yamanaka Atsushi、Yato Keigo、Yoshii Kentaro、Watashi Koichi、Aizaki Hideki、Konishi Eiji、Takasaki Tomohiko、Kato Takanobu、Muramatsu Masamichi、Wakita Takaji、Suzuki Ryosuke
  • Journal Title: Scientific Reports Volume: 8 Pages: －
  • DOI: 10.1038/s41598-018-34865-y
  • Peer Reviewed
• [Journal Article] The aryl hydrocarbon receptor?cytochrome P450 1A1 pathway controls lipid accumulation and enhances the permissiveness for hepatitis C virus assembly (2018)
  • Author(s): Ohashi Hirofumi、Nishioka Kazane、Nakajima Syo、Kim Sulyi、Suzuki Ryosuke、Aizaki Hideki、Fukasawa Masayoshi、Kamisuki Shinji、Sugawara Fumio、Ohtani Naoko、Muramatsu Masamichi、Wakita Takaji、Watashi Koichi
  • Journal Title: Journal of Biological Chemistry Volume: 293 Pages: 19559～19571
  • DOI: 10.1074/jbc.RA118.005033
  • Peer Reviewed
• [Journal Article] A new strategy to identify hepatitis B virus entry inhibitors by AlphaScreen technology targeting the envelope-receptor interaction (2018)
  • Author(s): Saso Wakana、Tsukuda Senko、Ohashi Hirofumi、Fukano Kento、Morishita Ryo、Matsunaga Satoko、Ohki Mio、Ryo Akihide、Park Sam-Yong、Suzuki Ryosuke、Aizaki Hideki、Muramatsu Masamichi、Sureau Camille、Wakita Takaji、Matano Tetsuro、Watashi Koichi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 501 Pages: 374～379
  • DOI: 10.1016/j.bbrc.2018.04.187
  • Peer Reviewed
• [Journal Article] Chemical array system, a platform to identify novel hepatitis B virus entry inhibitors targeting sodium taurocholate cotransporting polypeptide (2018)
  • Author(s): Kaneko M, Futamura Y, Tsukuda S, Kondoh Y, Sekine T, Hirano H, Fukano K, Ohashi H, Saso W, Morishita R, Matsunaga S, Kawai F, Ryo A, Park SY, Suzuki R, Aizaki H, Ohtani N, Sureau C, Wakita T, Osada H, Watashi K.
  • Journal Title: Scientific Reports Volume: 8 Pages: －
  • DOI: 10.1038/s41598-018-20987-w
  • Peer Reviewed
• [Presentation] Alyl hydrocarbon receptor-cytochrome p450 1a1 pathway regulates hepatic lipid biosynthesis to maximize Hepatitis C virus production. (2018)
  • Author(s): Hirofumi Ohashi, Syo Nakajima, Sulyl Kim, Ryosuke Suzuki, Hideki Aizaki, Masayoshi Fukasawa, Shinji Kamisuki, Fumio Sugawara, Naoko Ohtani, Masamichi Muramatsu, Takaji Wakita, Koichi Watashi.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Construction of chimeric reporter HCV with efficient production capacity. (2018)
  • Author(s): Rika Sato, Noriyuki Watanabe, Hussein Aly, Madoka Koyanagi, Yutaka Arimura, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses
  • Int'l Joint Research
• [Presentation] Establishment of infectious genotype 4a HCVcc. (2018)
  • Author(s): Noriyuki Watanabe, Takaya Suzuki, Tomoko Date, Su Su Hmwe, Hussein Aly, Hideki Aizaki, Masaya, Sugiyama, Masashi Mizokami, Mohamed El Kassas, Ashraf Tabll, William Delaney, Guofeng Cheng, Masamichi Muramatsu, Takaji Wakita.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Antiviral activity of phospholipase A2 group V (PLA2G5) against HCV. (2018)
  • Author(s): Francesc Puig-Basagoiti, Masayoshi Fukasawa, Ryosuke Suzuki, Koichi Watashi, Masamichi Muramatsu, Takaji Wakita, Hideki Aizaki.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Flavivirus subviral particles-based DNA vaccine induces neutralizing antibodies against HCV. (2018)
  • Author(s): Keigo Yato, Mami Matsuda, Noriyuki Watanabe, Shogo Nakajima, Akira Fujimoto, Koichi Watashi, Hideki Aizaki, Takanobu Kato, Koji Tamura, Masamichi Muramatsu, Takaji Wakita, Ryosuke Suzuki.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Identification of Alyl hydrocarbon receptor ligands inhibiting the lipid accumulation and hepatitis C virus production. (2018)
  • Author(s): Kazane Nishioka, Hirofumi Ohashi, Ryosuke Suzuki, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita, Koichi Watashi.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Abnormal hepatocellular organelles remain to be observed in sustained virological response (SVR) patients. (2018)
  • Author(s): Haruyo Aoyagi, Hiroko Iijima, Xin Zheng, Yu Ting Kao, Mami Matsuda, Koichi Watashi, Ryosuke Suzuki, Takahiro Masaki, Noritomo Shimada, Keizo Kato, Akihito Tsubota, Kenjiro Wake, Takaji Wakita, Hideki Aizaki.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Abnormal hepatocellular organelles remain to be observed in sustained virological response patients. (2018)
  • Author(s): Haruyo Aoyagi, et. al.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Human hepatic stellate cells are permissive for hepatitis C virus infection/replication and play important roles in fibrosis. (2018)
  • Author(s): Xin Zheng, et al.
  • Organizer: 25th International Symposium on Hepatitis C Virus and Related Viruses.
  • Int'l Joint Research
• [Presentation] Epidemiology Study of Acute Hepatitis B and C in Japan, from April 1999 (2018)
  • Author(s): Xin Zheng, Haruyo Aoyagi, Gewaid E. Hossam, Takuma Zaitsu, Francesc Puig-Basagoiti, Yu Ting Kao, Koichi Watashi, Ryosuke Suzuki, Takuri Takahashi, Tomimasa Sunagawa, Kazunori Oishi, Takaji Wakita, Hideki Aizaki.
  • Organizer: 6th JAPAN-TAIWAN-KOREA HBV Research Symposium 2018
• [Presentation] HCVに対する抗ウイルス治療後、SVR後の肝細胞の超微細構造の変化. (2018)
  • Author(s): 青柳東代, 飯島尋子, 松田麻未, 渡士幸一, 鈴木亮介, 政木隆博, 酒巻有里子, 市野瀬志津子, 坪田昭人, 和氣健二郎, 脇田隆字, 相崎英樹.
  • Organizer: 第２６回抗ウイルス療法学会総会
• [Presentation] Hepatitis C virus infection triggers the transavtivation of alyl hydrocarbon receptor to rearrange hepatic lipid biosynthesis. (2018)
  • Author(s): Hirofumi Ohashi, Syo Nakajima, Sulyl Kim, Ryosuke Suzuki, Hideki Aizaki, Masayoshi Fukasawa, Shinji Kamisuki, Fumio Sugawara, Naoko Ohtani, Masamichi Muramatsu, Takaji Wakita, Koichi Watashi.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Development and use of chronological and real-time monitoring system of hepatitis C virus RNA replication. (2018)
  • Author(s): Xiaoyu Li, Masahiko Ito, Kenji Nakashima, Haruyo Aoyagi, Hideki Aizaki, Tetsuro Suzuki.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] HCV genome and abnormal organelles in the liver after elimination of hepatitis C virus by drug treatment. (2018)
  • Author(s): Hideki Aizaki, Haruyo Aoyagi, Hiroko Iijima, Xin Zheng, Mami Matsuda, Koichi Watashi, Ryosuke Suzuki, Takahiro Masaki, Noritomo Shimada, Keizo Kato, Akihito Tsubota, Takeshi Saito, Kazuhiko Hayashi, Masaru Enomoto, Ayako Mimata, Yuriko Sakamaki, Shizuko Ichinose, Kenjiro Wake, Masamichi Muramatsu, Takaji Wakita.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Identification of Alyl hydrocarbon receptor ligands inhibiting the lipid accumulation and hepatitis C virus production. (2018)
  • Author(s): Kazane Nishioka, Hirofumi Ohashi, Ryosuke Suzuki, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita, Koichi Watashi.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Construction of chimeric reporter HCV efficient production capacity. (2018)
  • Author(s): Rika Sato, Noriyuki Watanabe, Hussein Aly, Madoka Koyanagi, Yutaka Arimura, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Establishment of infectious genotype 4a HCVcc. (2018)
  • Author(s): Noriyuki Watanabe, Takaya Suzuki, Tomoko Date, Su Su Hmwe, Hussein Aly, Masaya Sugiyama, Masashi Mizokami, Hideki Aizaki, Masamichi Muramatsu, Takaji Wakita.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Book] C型肝炎ウイルスの遺伝子構造と抗ウイルス薬の作用機序は？ (2018)
  • Author(s): 52.相崎英樹
  • Total Pages: 1
  • Publisher: 日本医事新報