title: N6-Methyladenosine in Flaviviridae Viral RNA Genomes Regulates Infection.
abstract: The RNA modification N6-methyladenosine (m6A) post-transcriptionally regulates RNA function. The cellular machinery that controls m6A includes methyltransferases and demethylases that add or remove this modification, as well as m6A-binding YTHDF proteins that promote the translation or degradation of m6A-modified mRNA. We demonstrate that m6A modulates infection by hepatitis C virus (HCV). Depletion of m6A methyltransferases or an m6A demethylase, respectively, increases or decreases infectious HCV particle production. During HCV infection, YTHDF proteins relocalize to lipid droplets, sites of viral assembly, and their depletion increases infectious viral particles. We further mapped m6A sites across the HCV genome and determined that inactivating m6A in one viral genomic region increases viral titer without affecting RNA replication. Additional mapping of m6A on the RNA genomes of other Flaviviridae, including dengue, Zika, yellow fever, and West Nile virus, identifies conserved regions modified by m6A. Altogether, this work identifies m6A as a conserved regulatory mark across Flaviviridae genomes.
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Nandan S
authors: NS
authors: Gokhale
authors: Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10021, USA; Tri-Institutional Program in Computational Biology and Medicine, New York City, NY 10065, USA.
authors: Alexa B R
authors: ABR
authors: McIntyre
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Michael J
authors: MJ
authors: McFadden
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Allison E
authors: AE
authors: Roder
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Edward M
authors: EM
authors: Kennedy
authors: Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10021, USA.
authors: Jorge A
authors: JA
authors: Gandara
authors: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors: Sharon E
authors: SE
authors: Hopcraft
authors: Division of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Vaccine Center, Yerkes National Primate Research Center, Atlanta, GA 30329, USA.
authors: Kendra M
authors: KM
authors: Quicke
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Christine
authors: C
authors: Vazquez
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
authors: Jason
authors: J
authors: Willer
authors: Duke Molecular Physiology Institute, Duke University, Durham NC 27701, USA.
authors: Olga R
authors: OR
authors: Ilkayeva
authors: Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
authors: Brittany A
authors: BA
authors: Law
authors: Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
authors: Christopher L
authors: CL
authors: Holley
authors: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, USA; Programme in Emerging Infectious Disease, Duke-NUS Medical School, Singapore 169857, Singapore.
authors: Mariano A
authors: MA
authors: Garcia-Blanco
authors: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors: Matthew J
authors: MJ
authors: Evans
authors: Division of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, USA; Emory Vaccine Center, Yerkes National Primate Research Center, Atlanta, GA 30329, USA.
authors: Mehul S
authors: MS
authors: Suthar
authors: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors: Shelton S
authors: SS
authors: Bradrick
authors: Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10021, USA; The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY 10021, USA; The Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York, NY 10021, USA. Electronic address: chm2042@med.cornell.edu.
authors: Christopher E
authors: CE
authors: Mason
authors: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA; Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA. Electronic address: stacy.horner@duke.edu.
authors: Stacy M
authors: SM
authors: Horner
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism; D018067:Flaviviridae / Q000235:genetics* / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D054884:Host-Pathogen Interactions; D008780:Methyltransferases / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D019562:Viral Load; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; C010223:N-methyladenosine; D010089:Oxidoreductases, N-Demethylating; D008780:Methyltransferases; D000241:Adenosine
keywords: Flaviviridae; HCV; N6-methyladenosine; RNA-modifications; West Nile; Zika; dengue; m(6)A; viral particle production; yellow fever