title: The primary structure and expression of the second open reading frame of the polymerase gene of the coronavirus MHV-A59; a highly conserved polymerase is expressed by an efficient ribosomal frameshifting mechanism.
abstract: Sequence analysis of a substantial part of the polymerase gene of the murine coronavirus MHV-A59 revealed the 3' end of an open reading frame (ORF1a) overlapping with a large ORF (ORF1b; 2733 amino acids) which covers the 3' half of the polymerase gene. The expression of ORF1b occurs by a ribosomal frameshifting mechanism since the ORF1a/ORF1b overlapping nucleotide sequence is capable of inducing ribosomal frameshifting in vitro as well as in vivo. A stem-loop structure and a pseudoknot are predicted in the nucleotide sequence involved in ribosomal frameshifting. Comparison of the predicted amino acid sequence of MHV ORF1b with the amino acid sequence deduced from the corresponding gene of the avian coronavirus IBV demonstrated that in contrast to the other viral genes this ORF is extremely conserved. Detailed analysis of the predicted amino acid sequence revealed sequence elements which are conserved in many DNA and RNA polymerases.
authors.affiliation: Department of Infectious Diseases and Immunology, State University of Utrecht, The Netherlands.
authors.forename: P J
authors.initials: PJ
authors.lastname: Bredenbeek
authors.forename: C J
authors.initials: CJ
authors.lastname: Pachuk
authors.forename: A F
authors.initials: AF
authors.lastname: Noten
authors.forename: J
authors.initials: J
authors.lastname: Charité
authors.forename: W
authors.initials: W
authors.lastname: Luytjes
authors.forename: S R
authors.initials: SR
authors.lastname: Weiss
authors.forename: W J
authors.initials: WJ
authors.lastname: Spaan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D015870:Gene Expression; D005814:Genes, Viral; D051379:Mice; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000201:enzymology / Q000235:genetics*; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D012270:Ribosomes / Q000378:metabolism
chemical_list: D015345:Oligonucleotide Probes; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases