title: Unique Requirement for ESCRT Factors in Flavivirus Particle Formation on the Endoplasmic Reticulum.
abstract: Flavivirus infection induces endoplasmic reticulum (ER) membrane rearrangements to generate a compartment for replication of the viral genome and assembly of viral particles. Using quantitative mass spectrometry, we identified several ESCRT (endosomal sorting complex required for transport) proteins that are recruited to sites of virus replication on the ER. Systematic small interfering RNA (siRNA) screening revealed that release of both dengue virus and Japanese encephalitis virus was dramatically decreased by single depletion of TSG101 or co-depletion of specific combinations of ESCRT-III proteins, resulting in ≥1,000-fold titer reductions. By contrast, release was unaffected by depletion of some core ESCRTs, including VPS4. Reintroduction of ESCRT proteins to siRNA-depleted cells revealed interactions among ESCRT proteins that are crucial for flavivirus budding. Electron-microscopy studies revealed that the CHMP2 and CHMP4 proteins function directly in membrane deformation at the ER. Thus, a unique and specific subset of ESCRT contributes to ER membrane biogenesis during flavivirus infection.
authors.affiliation: Laboratory of Viral Infection, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Tabata
authors.affiliation: Laboratory of Viral Infection, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Arimoto
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki 036-8561, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Arakawa
authors.affiliation: Faculty of Bioscience, Nagahama Institute of Bio-science and Technology, Nagahama 526-0829, Japan.
authors.forename: Atsuki
authors.initials: A
authors.lastname: Nara
authors.affiliation: Core Instrumentation Facility, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Saito
authors.affiliation: Core Instrumentation Facility, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Omori
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki 036-8561, Japan.
authors.forename: Arisa
authors.initials: A
authors.lastname: Arai
authors.affiliation: Department of Microbiology, Dokkyo Medical University School of Medicine, Tochigi 321-0293 Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ishikawa
authors.affiliation: BIKEN Endowed Department of Dengue Vaccine Development, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan; BIKEN Endowed Department of Dengue Vaccine Development, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Eiji
authors.initials: E
authors.lastname: Konishi
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Matsuura
authors.affiliation: Laboratory of Viral Infection, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan; Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki 036-8561, Japan. Electronic address: moritae@hirosaki-u.ac.jp.
authors.forename: Eiji
authors.initials: E
authors.lastname: Morita
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003412:Cricetulus; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D003716:Dengue Virus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics* / Q000821:virology; D056827:Endosomal Sorting Complexes Required for Transport / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism* / Q000821:virology; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D025262:Vacuolar Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells; D014771:Virion / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics; D000099177:Tsg101 Protein
chemical_list: D000074183:ATPases Associated with Diverse Cellular Activities; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D034741:RNA, Small Interfering; D014157:Transcription Factors; D025262:Vacuolar Proton-Translocating ATPases; D000099177:Tsg101 Protein; C507674:CHMP2A protein, human; C481529:CHMP4A protein, human; C490305:VPS4A protein, human
keywords: CHMP2; CHMP4; ESCRT; TSG101; flavivirus