title: A cytoplasmic RNA virus generates functional viral small RNAs and regulates viral IRES activity in mammalian cells.
abstract: The roles of virus-derived small RNAs (vsRNAs) have been studied in plants and insects. However, the generation and function of small RNAs from cytoplasmic RNA viruses in mammalian cells remain unexplored. This study describes four vsRNAs that were detected in enterovirus 71-infected cells using next-generation sequencing and northern blots. Viral infection produced substantial levels (>10(5) copy numbers per cell) of vsRNA1, one of the four vsRNAs. We also demonstrated that Dicer is involved in vsRNA1 generation in infected cells. vsRNA1 overexpression inhibited viral translation and internal ribosomal entry site (IRES) activity in infected cells. Conversely, blocking vsRNA1 enhanced viral yield and viral protein synthesis. We also present evidence that vsRNA1 targets stem-loop II of the viral 5' untranslated region and inhibits the activity of the IRES through this sequence-specific targeting. Our study demonstrates the ability of a cytoplasmic RNA virus to generate functional vsRNA in mammalian cells. In addition, we also demonstrate a potential novel mechanism for a positive-stranded RNA virus to regulate viral translation: generating a vsRNA that targets the IRES.
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Center for Molecular and Clinical Immunology, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Kuo-Feng
authors.initials: KF
authors.lastname: Weng
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Chuan-Tien
authors.initials: CT
authors.lastname: Hung
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Po-Ting
authors.initials: PT
authors.lastname: Hsieh
authors.affiliation: Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ, USA.
authors.forename: Mei-Ling
authors.initials: ML
authors.lastname: Li
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Department of Computer Science and Information Engineering, College of Engineering, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Guang-Wu
authors.initials: GW
authors.lastname: Chen
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Yu-An
authors.initials: YA
authors.lastname: Kung
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Peng-Nien
authors.initials: PN
authors.lastname: Huang
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Rei-Lin
authors.initials: RL
authors.lastname: Kuo
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Li-Lien
authors.initials: LL
authors.lastname: Chen
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taiwan.
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Lin
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Robert Yung-Liang
authors.initials: RY
authors.lastname: Wang
authors.affiliation: Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Shu-Jen
authors.initials: SJ
authors.lastname: Chen
authors.affiliation: Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Petrus
authors.initials: P
authors.lastname: Tang
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Jim-Tong
authors.initials: JT
authors.lastname: Horng
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Hsing-I
authors.initials: HI
authors.lastname: Huang
authors.affiliation: Center of Infectious Disease and Signaling Research and Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Jen-Ren
authors.initials: JR
authors.lastname: Wang
authors.affiliation: Center for Molecular and Clinical Immunology, College of Medicine, Chang Gung University, Taoyuan, Taiwan Health Sciences Research Institute and School of Natural Sciences, University of California, Merced, CA, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Ojcius
authors.affiliation: Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ, USA.
authors.forename: Gary
authors.initials: G
authors.lastname: Brewer
authors.affiliation: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan Clinical Virology Laboratory, Chang Gung Memorial Hospital, Tao-yuan, Taiwan srshih@mail.cgu.edu.tw.
authors.forename: Shin-Ru
authors.initials: SR
authors.lastname: Shih
mesh_terms: D020121:5' Untranslated Regions; D045744:Cell Line, Tumor; D015536:Down-Regulation; D029821:Enterovirus A, Human / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D006801:Humans; D014176:Protein Biosynthesis; D058727:RNA, Small Untranslated / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D043244:Ribonuclease III / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis
chemical_list: D020121:5' Untranslated Regions; D058727:RNA, Small Untranslated; D012367:RNA, Viral; D014764:Viral Proteins; D043244:Ribonuclease III