title: UVRAG is required for virus entry through combinatorial interaction with the class C-Vps complex and SNAREs.
abstract: Enveloped viruses exploit the endomembrane system to enter host cells. Through a cascade of membrane-trafficking events, virus-bearing vesicles fuse with acidic endosomes and/or lysosomes mediated by SNAREs triggering viral fusion. However, the molecular mechanisms underlying this process remain elusive. Here, we found that UV-radiation resistance-associated gene (UVRAG), an autophagic tumor suppressor, is required for the entry of the prototypic negative-strand RNA virus, including influenza A virus and vesicular stomatitis virus, by a mechanism independent of IFN and autophagy. UVRAG mediates viral endocytic transport and membrane penetration through interactions with the class C vacuolar protein sorting (C-Vps) tethering complex and endosomal glutamine-containing SNAREs [syntaxin 7 (STX7), STX8, and vesicle transport through t-SNARE homolog 1B (Vti1b)], leading to the assembly of a fusogenic trans-SNARE complex involving vesicle-associated membrane protein (VAMP8), but not VAMP7. Indeed, UVRAG stimulates VAMP8 translocation to virus-bearing endosomes. Inhibition of VAMP8, but not VAMP7, significantly reduces viral entry. Our data indicate that UVRAG, in concert with C-Vps, regulates viral entry by assembling a specific fusogenic SNARE complex. Thus, UVRAG governs downstream viral entry, highlighting an important pathway capable of potential antiviral therapeutics.
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033.
authors.forename: Sara Dolatshahi
authors.initials: SD
authors.lastname: Pirooz
authors.forename: Shanshan
authors.initials: S
authors.lastname: He
authors.forename: Tian
authors.initials: T
authors.lastname: Zhang
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhao
authors.forename: Soohwan
authors.initials: S
authors.lastname: Oh
authors.forename: Douglas
authors.initials: D
authors.lastname: O'Connell
authors.forename: Payam
authors.initials: P
authors.lastname: Khalilzadeh
authors.forename: Samad
authors.initials: S
authors.lastname: Amini-Bavil-Olyaee
authors.forename: Michael
authors.initials: M
authors.lastname: Farzan
authors.forename: Chengyu
authors.initials: C
authors.lastname: Liang
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D006224:Cricetinae; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D009980:Influenza A virus / Q000502:physiology; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000235:genetics; D050683:R-SNARE Proteins / Q000378:metabolism*; D034622:RNA Interference; D012328:RNA Viruses / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D014709:Vero Cells; D033921:Vesicular Transport Proteins / Q000378:metabolism*; D018116:Vesiculovirus / Q000502:physiology; D053586:Virus Internalization
chemical_list: D050683:R-SNARE Proteins; D034741:RNA, Small Interfering; D025521:Tumor Suppressor Proteins; C408122:UVRAG protein, human; C112938:VAMP8 protein, human; D033921:Vesicular Transport Proteins; D049452:Green Fluorescent Proteins
keywords: VSV; endocytic trafficking; influenza virus