title: Characterization of RyDEN (C19orf66) as an Interferon-Stimulated Cellular Inhibitor against Dengue Virus Replication.
abstract: Dengue virus (DENV) is one of the most important arthropod-borne pathogens that cause life-threatening diseases in humans. However, no vaccine or specific antiviral is available for dengue. As seen in other RNA viruses, the innate immune system plays a key role in controlling DENV infection and disease outcome. Although the interferon (IFN) response, which is central to host protective immunity, has been reported to limit DENV replication, the molecular details of how DENV infection is modulated by IFN treatment are elusive. In this study, by employing a gain-of-function screen using a type I IFN-treated cell-derived cDNA library, we identified a previously uncharacterized gene, C19orf66, as an IFN-stimulated gene (ISG) that inhibits DENV replication, which we named Repressor of yield of DENV (RyDEN). Overexpression and gene knockdown experiments revealed that expression of RyDEN confers resistance to all serotypes of DENV in human cells. RyDEN expression also limited the replication of hepatitis C virus, Kunjin virus, Chikungunya virus, herpes simplex virus type 1, and human adenovirus. Importantly, RyDEN was considered to be a crucial effector molecule in the IFN-mediated anti-DENV response. When affinity purification-mass spectrometry analysis was performed, RyDEN was revealed to form a complex with cellular mRNA-binding proteins, poly(A)-binding protein cytoplasmic 1 (PABPC1), and La motif-related protein 1 (LARP1). Interestingly, PABPC1 and LARP1 were found to be positive modulators of DENV replication. Since RyDEN influenced intracellular events on DENV replication and, suppression of protein synthesis from DENV-based reporter construct RNA was also observed in RyDEN-expressing cells, our data suggest that RyDEN is likely to interfere with the translation of DENV via interaction with viral RNA and cellular mRNA-binding proteins, resulting in the inhibition of virus replication in infected cells.
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.|Emerging Infectious Disease Program, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Youichi
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Wei-Xin
authors.initials: WX
authors.lastname: Chin
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Qi'En
authors.initials: Q
authors.lastname: Han
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Koji
authors.initials: K
authors.lastname: Ichiyama
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Ching Hua
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Zhi Wen
authors.initials: ZW
authors.lastname: Eyo
authors.affiliation: Laboratory of Viral Pathogenesis, Institute for Virus Research, Kyoto University, Kyoto, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Ebina
authors.affiliation: Proteo-Science Center, Ehime University, Matsuyama, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Proteo-Science Center, Ehime University, Matsuyama, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Takahashi
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Beng Hui
authors.initials: BH
authors.lastname: Tan
authors.affiliation: Laboratory of Primate Model, Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Kyoto, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Hishiki
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Kenji
authors.initials: K
authors.lastname: Ohba
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Matsuyama
authors.affiliation: Laboratory of Viral Pathogenesis, Institute for Virus Research, Kyoto University, Kyoto, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Koyanagi
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Yee-Joo
authors.initials: YJ
authors.lastname: Tan
authors.affiliation: Proteo-Science Center, Ehime University, Matsuyama, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Sawasaki
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Justin Jang Hann
authors.initials: JJ
authors.lastname: Chu
authors.affiliation: Emerging Infectious Disease Program, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Subhash G
authors.initials: SG
authors.lastname: Vasudevan
authors.affiliation: Department of Microbiology and Infection Control, Osaka Medical College, Takatsuki, Japan.
authors.forename: Kouichi
authors.initials: K
authors.lastname: Sano
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Naoki
authors.initials: N
authors.lastname: Yamamoto
mesh_terms: D002460:Cell Line; D003715:Dengue / Q000276:immunology*; D003716:Dengue Virus / Q000254:growth & development / Q000502:physiology*; D055785:Gene Knockdown Techniques; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D007372:Interferons / Q000276:immunology*; D013058:Mass Spectrometry; D016133:Polymerase Chain Reaction; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication / Q000276:immunology*
chemical_list: D014764:Viral Proteins; D007372:Interferons