An 'elaborated' pseudoknot is required for high frequency frameshifting during translation of HCV 229E polymerase mRNA.
The RNA polymerase gene (gene 1) of the human coronavirus 229E is approximately 20 kb in length and is located at the 5' end of the positive-strand genomic RNA. The coding sequence of gene 1 is divided into two large open reading frames, ORF1a and ORF1b, that overlap by 43 nucleotides. In the region of the ORF1a/ORF1b overlap, the genomic RNA displays two elements that are known to mediate (-1) ribosomal frameshifting. These are the slippery sequence, UUUAAAC, and a 3' pseudoknot structure. By introducing site-specific mutations into synthetic mRNAs, we have analysed the predicted structure of the HCV 229E pseudoknot and shown that besides the well-known stem structures, S1 and S2, a third stem structure, S3, is required for a high frequency of frameshifting. The requirement for an S3 stem is independent of the length of loop 2.
Herold
J
J
Institute of Virology, University of Würzburg, Germany.
Siddell
S G
SG
D000595:Amino Acid Sequence; D001482:Base Composition; D001483:Base Sequence; D017934:Coronavirus / Q000235:genetics*; D028941:Coronavirus 229E, Human; D004247:DNA; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D016368:Frameshift Mutation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics*
D012367:RNA, Viral; D004247:DNA; D012321:DNA-Directed RNA Polymerases