PMID:17704133
| Citation |
Léger, M, Dulude, D, Steinberg, SV and Brakier-Gingras, L (2007) The three transfer RNAs occupying the A, P and E sites on the ribosome are involved in viral programmed -1 ribosomal frameshift. Nucleic Acids Res. 35:5581-92 |
|---|---|
| Abstract |
The -1 programmed ribosomal frameshifts (PRF), which are used by many viruses, occur at a heptanucleotide slippery sequence and are currently thought to involve the tRNAs interacting with the ribosomal P- and A-site codons. We investigated here whether the tRNA occupying the ribosomal E site that precedes a slippery site influences -1 PRF. Using the human immunodeficiency virus type 1 (HIV-1) frameshift region, we found that mutating the E-site codon altered the -1 PRF efficiency. When the HIV-1 slippery sequence was replaced with other viral slippery sequences, mutating the E-site codon also altered the -1 PRF efficiency. Because HIV-1 -1 PRF can be recapitulated in bacteria, we used a bacterial ribosome system to select, by random mutagenesis, 16S ribosomal RNA (rRNA) mutations that modify the expression of a reporter requiring HIV-1 -1 PRF. Three mutants were isolated, which are located in helices 21 and 22 of 16S rRNA, a region involved in translocation and E-site tRNA binding. We propose a novel model where -1 PRF is triggered by an incomplete translocation and depends not only on the tRNAs interacting with the P- and A-site codons, but also on the tRNA occupying the E site. |
| Links |
PubMed PMC2018615 Online version:10.1093/nar/gkm578 |
| Keywords |
Cell Line; Codon/chemistry; Frameshifting, Ribosomal; Genes, Reporter; HIV-1/genetics; Humans; Models, Genetic; Mutation; Nucleotides/chemistry; RNA, Messenger/chemistry; RNA, Ribosomal, 16S/chemistry; RNA, Transfer/metabolism; RNA, Viral/chemistry; Ribosomes/chemistry; Ribosomes/metabolism |
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