title: HCV IRES manipulates the ribosome to promote the switch from translation initiation to elongation.
abstract: The internal ribosome entry site (IRES) of the hepatitis C virus (HCV) drives noncanonical initiation of protein synthesis necessary for viral replication. Functional studies of the HCV IRES have focused on 80S ribosome formation but have not explored its role after the 80S ribosome is poised at the start codon. Here, we report that mutations of an IRES domain that docks in the 40S subunit's decoding groove cause only a local perturbation in IRES structure and result in conformational changes in the IRES-rabbit 40S subunit complex. Functionally, the mutations decrease IRES activity by inhibiting the first ribosomal translocation event, and modeling results suggest that this effect occurs through an interaction with a single ribosomal protein. The ability of the HCV IRES to manipulate the ribosome provides insight into how the ribosome's structure and function can be altered by bound RNAs, including those derived from cellular invaders.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver School of Medicine, Aurora, Colorado, USA.
authors.forename: Megan E
authors.initials: ME
authors.lastname: Filbin
authors.forename: Breanna S
authors.initials: BS
authors.lastname: Vollmar
authors.forename: Dan
authors.initials: D
authors.lastname: Shi
authors.forename: Tamir
authors.initials: T
authors.lastname: Gonen
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Kieft
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D020285:Cryoelectron Microscopy; D017931:DNA Primers / Q000235:genetics; D016174:Hepacivirus / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008156:Luciferases; D009682:Magnetic Resonance Spectroscopy; D008854:Microscopy, Electron; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D010441:Peptide Chain Elongation, Translational / Q000502:physiology*; D010442:Peptide Chain Initiation, Translational / Q000502:physiology*; D010957:Plasmids / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D054682:Ribosome Subunits, Small, Eukaryotic / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral; D008156:Luciferases