Nucleic Acids Research Advance Access originally published online on August 17, 2007
Nucleic Acids Research 2007 35(16):5581-5592; doi:10.1093/nar/gkm578
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Nucleic Acids Research, 2007, Vol. 35, No. 16 5581-5592
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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The three transfer RNAs occupying the A, P and E sites on the ribosome are involved in viral programmed -1 ribosomal frameshift
Département de Biochimie, Université de Montréal, Montréal, Québec, Canada, H3T 1J4
*To whom correspondence should be addressed. Tel: 514 343 6316; Fax: 514 343 2210; Email: lea.brakier.gingras{at}umontreal.ca
Received May 30, 2007. Revised June 28, 2007. Accepted July 13, 2007.
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.
Present address: Dominic Dulude, Centre de recherche, Hôpital Ste-Justine, 3175, chemin de la Côte-Ste-Catherine, Montréal, Québec, Canada H3T 1C5
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