title: Ribosomal proteins mediate the hepatitis C virus IRES-HeLa 40S interaction.
abstract: Translation of the hepatitis C virus genomic RNA is mediated by an internal ribosome entry site (IRES). The 330-nt IRES RNA forms a binary complex with the small 40S ribosomal subunit as a first step in translation initiation. Here chemical probing and 4-thiouridine-mediated crosslinking are used to characterize the interaction of the HCV IRES with the HeLa 40S subunit. No IRES-18S rRNA contacts were detected, but several specific crosslinks to 40S ribosomal proteins were observed. The identity of the crosslinked proteins agrees well with available structural information and provides new insights into HCV IRES function. The protein-rich surface of the 40S subunit thus mediates the IRES-ribosome interaction.
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, California 94305-5126, USA.
authors.forename: Geoff A
authors.initials: GA
authors.lastname: Otto
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lukavsky
authors.forename: Alissa M
authors.initials: AM
authors.lastname: Lancaster
authors.forename: Peter
authors.initials: P
authors.lastname: Sarnow
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Puglisi
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003432:Cross-Linking Reagents; D015180:Electrophoresis, Gel, Two-Dimensional; D016679:Genome, Viral; D006367:HeLa Cells; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010442:Peptide Chain Initiation, Translational; D021122:Protein Subunits; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013891:Thiouridine
chemical_list: D003432:Cross-Linking Reagents; D021122:Protein Subunits; D012367:RNA, Viral; D012269:Ribosomal Proteins; C029537:ribosomal protein S10; D013891:Thiouridine