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Nucleic Acids Research Advance Access published online on October 9, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp816
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

Nucleotide modifications in three functionally important regions of the Saccharomyces cerevisiae ribosome affect translation accuracy

Agnès Baudin-Baillieu1,2,*, Céline Fabret1,2, Xue-hai Liang3, Dorota Piekna-Przybylska3, Maurille J. Fournier3 and Jean-Pierre Rousset1,4

1IGM, CNRS, UMR 8621, Orsay, F 91405, 2Univ Pierre et Marie Curie, Paris, F 75006, France, 3Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA 01003, USA and 4Univ Paris-Sud, Orsay, F 91405, France

*To whom correspondence should be addressed. Email: abaillieu{at}igmors.u-psud.fr

Received June 18, 2009. Revised September 11, 2009. Accepted September 15, 2009.

Important regions of rRNA are rich in nucleotide modifications that can have strong effects on ribosome biogenesis and translation efficiency. Here, we examine the influence of pseudouridylation and 2'-O-methylation on translation accuracy in yeast, by deleting the corresponding guide snoRNAs. The regions analyzed were: the decoding centre (eight modifications), and two intersubunit bridge domains—the A-site finger and Helix 69 (six and five modifications). Results show that a number of modifications influence accuracy with effects ranging from 0.3- to 2.4-fold of wild-type activity. Blocking subsets of modifications, especially from the decoding region, impairs stop codon termination and reading frame maintenance. Unexpectedly, several Helix 69 mutants possess ribosomes with increased fidelity. Consistent with strong positional and synergistic effects is the finding that single deletions can have a more pronounced phenotype than multiple deficiencies in the same region. Altogether, the results demonstrate that rRNA modifications have significant roles in translation accuracy.


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