Nucleic Acids Research, Vol 26, Issue 7 1576-1587, Copyright © 1998 by Oxford University Press
J Cavaille and JP Bachellerie
Eukaryotic rRNAs contain a large number of ribose-methylated nucleotides of
elusive function which are confined to the universally conserved rRNA
domains. Ribose methylation of these nucleotides is directed by a large
family of small trans -acting guide RNAs, called box C/D antisense snoRNAs.
Each snoRNA targets precisely one of the nucleotides to be methylated
within the pre-rRNA sequence, through transient formation of a 10-21 bp
regular RNA duplex around the modification site. In this study we have
analyzed how different features of the double-stranded RNA guide structure
affect the extent of site-specific ribose methylation, by co-expressing an
appropriate RNA substrate and its cognate tailored snoRNA guide in
transfected mouse cells. We show that an increased GC content of the duplex
can make up for the inhibitory effects of a helix truncation or for the
presence of helix irregularities such as a mismatched pair or a bulge
nucleotide. However, some helix irregularities dramatically inhibit the
reaction and are not offset by further stabilization of the duplex.
Overall, the RNA duplex tolerates a much larger degree of irregularity than
anticipated, even in the immediate vicinity of the methylation site, which
offers new prospects in the search for additional snoRNA guides.
Accordingly, a few snoRNA-like sequences of uncertain status detected in
the yeast Saccharomyces cerevisiae genome now appear as likely bona fide
ribose methylation guides.
ARTICLES
SnoRNA-guided ribose methylation of rRNA: structural features of the guide RNA duplex influencing the extent of the reaction
Laboratoire de Biologie Moleculaire Eucaryote du CNRS, Universite Paul- Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex, France.
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