title: TGEV corona virus ORF4 encodes a membrane protein that is incorporated into virions.
abstract: The coding potential of the open reading frame ORF4 (82 amino acids) of transmissible gastroenteritis virus (TGEV) has been confirmed by expression using a baculovirus vector. Five monoclonal antibodies (MAbs) raised against the 10K recombinant product immunoprecipitated a polypeptide of a similar size in TGEV-infected cells. Immunofluorescence assays performed both on insect and mammalian cells revealed that ORF4 was a membrane-associated protein, a finding consistent with the prediction of a membrane-spanning segment in ORF4 sequence. Two epitopes were localized within the last 21 C-terminal residues of the sequence through peptide scanning and analysis of the reactivity of a truncated ORF4 recombinant protein. Since the relevant MAbs were found to induce a cell surface fluorescence, these data suggest that ORF4 may be an integral membrane protein having a Cexo-Nendo orientation. Anti-ORF4 MAbs were also used to show that ORF4 polypeptide may be detected in TGEV virion preparations, with an estimated number of 20 molecules incorporated per particle. Comparison of amino acid sequence data provided strong evidence that other coronaviruses encode a polypeptide homologous to TGEV ORF4. Our results led us to propose that ORF4 represents a novel minor structural polypeptide, tentatively designated SM (small membrane protein).
authors.affiliation: Unité de Virologie et Immunologie Moléculaires, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: M
authors.initials: M
authors.lastname: Godet
authors.forename: R
authors.initials: R
authors.lastname: L'Haridon
authors.forename: J F
authors.initials: JF
authors.lastname: Vautherot
authors.forename: H
authors.initials: H
authors.lastname: Laude
mesh_terms: D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D016384:Consensus Sequence; D005814:Genes, Viral; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016366:Open Reading Frames; D010455:Peptides / Q000276:immunology; D016415:Sequence Alignment; D005760:Transmissible gastroenteritis virus / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics*; D014771:Virion / Q000737:chemistry
chemical_list: D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D014764:Viral Proteins; D015678:Viral Structural Proteins