The three transfer RNAs occupying the A, P and E sites on the ribosome are involved in viral programmed -1 ribosomal frameshift.
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.
Léger
Mélissa
M
Département de Biochimie, Université de Montréal, Montréal, Québec, Canada, H3T 1J4.
Dulude
Dominic
D
Steinberg
Sergey V
SV
Brakier-Gingras
Léa
L
D002460:Cell Line; D003062:Codon / Q000737:chemistry; D018965:Frameshifting, Ribosomal; D017930:Genes, Reporter; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008957:Models, Genetic; D009154:Mutation; D009711:Nucleotides / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry; D012336:RNA, Ribosomal, 16S / Q000737:chemistry; D012343:RNA, Transfer / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism
D003062:Codon; D009711:Nucleotides; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; D012367:RNA, Viral; D012343:RNA, Transfer
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