Stimulation of ribosomal frameshifting by antisense LNA

Programmed ribosomal frameshifting is a translational recoding mechanism commonly used by RNA viruses to express two or more proteins from a single mRNA at a fixed ratio. An essential element in this process is the presence of an RNA secondary structure, such as a pseudoknot or a hairpin, located do...

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Bibliographic Details
Published in:Nucleic Acids Research 2010-12, Vol.38 (22), p.8277-8283
Main Authors: Yu, Chien-Hung, Noteborn, Mathieu H.M, Olsthoorn, René C.L
Format: Article
Language:English
Subjects:
Antisense oligonucleotides
Data processing
Frameshifting, Ribosomal
mRNA
Nucleotide sequence
Oligodeoxyribonucleotides - chemistry
Oligonucleotides - chemistry
Oligonucleotides, Antisense - chemistry
Protein structure
RNA
RNA viruses
Secondary structure
Thermodynamics
Translation
Tunnels
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Summary:Programmed ribosomal frameshifting is a translational recoding mechanism commonly used by RNA viruses to express two or more proteins from a single mRNA at a fixed ratio. An essential element in this process is the presence of an RNA secondary structure, such as a pseudoknot or a hairpin, located downstream of the slippery sequence. Here, we have tested the efficiency of RNA oligonucleotides annealing downstream of the slippery sequence to induce frameshifting in vitro. Maximal frameshifting was observed with oligonucleotides of 12-18 nt. Antisense oligonucleotides bearing locked nucleid acid (LNA) modifications also proved to be efficient frameshift-stimulators in contrast to DNA oligonucleotides. The number, sequence and location of LNA bases in an otherwise DNA oligonucleotide have to be carefully manipulated to obtain optimal levels of frameshifting. Our data favor a model in which RNA stability at the entrance of the ribosomal tunnel is the major determinant of stimulating slippage rather than a specific three-dimensional structure of the stimulating RNA element.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/gkq650