Targeted ribose methylation of RNA in vivo directed by tailored antisense RNA guides

  title={Targeted ribose methylation of RNA in vivo directed by tailored antisense RNA guides},
  author={J{\'e}r{\^o}me Cavaill{\'e} and Monique Nicoloso and Jean Pierre Bachellerie},
EUKARYOTIC ribosomal RNAs are post-transcriptionally modified by methylation at the ribose sugar of specific nucleotides1. This takes place in the nucleolus and involves a family of small nucleolar RNAs (snoRNAs) with long regions (10–21 nucleotides) complementary to rRNA sequences spanning the methylation site2–4—a complementary snoRNA is required for methylation at a specific site5. Here we show that altering the sequence of the snoRNA is sufficient to change the specificity of methylation… 
Probing RNA in vivo with methylation guide small nucleolar RNAs.
A method is developed for constructing libraries of snoRNA genes that, in principle, can introduce methylation point mutations into any rRNA segment of interest and preliminary results are presented that show the feasibility of using this technology to probe a region of the yeast large subunit rRNA that includes the core of the peptidyltransferase center.
SnoRNA-guided ribose methylation of rRNA: structural features of the guide RNA duplex influencing the extent of the reaction.
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.
Small Nucleolar RNAs Guide the Ribose Methylations of Eukaryotic rRNAs
This chapter provides a progress report on developments that should set the stage for dissecting the rRNA-ribose methylation machinery and for deciphering the role of these modifications in ribosome
Interference probing of rRNA with snoRNPs: a novel approach for functional mapping of RNA in vivo.
It is reported that methylation snoRNPs can be targeted to many new sites in yeast rRNA, by providing the snoRNA with a novel guide sequence, and that in some cases growth and translation activity are strongly impaired.
Small nucleolar RNA‐guided post‐transcriptional modification of cellular RNAs
  • T. Kiss
  • Biology, Chemistry
    The EMBO journal
  • 2001
Post‐transcriptional modification of individual ribonucleotides, namely deamination of adenosines and cytidines, can also change the readout of mRNAs, and lack of ribosomal pseudouridines can reduce the growth rate or confer a selective disadvantage when it is competed against wild‐type ribosomes.
Targeted pre-mRNA modification for gene silencing and regulation
It is demonstrated that a Box C/D RNA can guide modification at the pre-mRNA branch point, thus silencing its expression and inducing cell death.
Structure and biogenesis of small nucleolar RNAs acting as guides for ribosomal RNA modification.
Information is reviewed about the biogenesis, structure and function of guide snoRNAs, which function as guides directing site-specific 2'-O-ribose methylation or pseudouridine formation in eukaryotic cells.
Profiling of RNA ribose methylation in Arabidopsis thaliana
The RiboMeth-seq data reveal the comprehensive 2′-O-methylation maps for Arabidopsis r RNAs and snRNAs and would facilitate study of their function and biosynthesis and the role of FIB1 or FIB2 in methylation.
RNA nucleotide methylation
This review outlines the different functions of methyl groups in RNA, including biophysical, biochemical and metabolic stabilization of RNA, quality control, resistance to antibiotics, mRNA reading frame maintenance, deciphering of normal and altered genetic code, selenocysteine incorporation, tRNA aminoacylation, ribotoxins, splicing, intracellular trafficking, immune response, and others.


A mammalian gene with introns instead of exons generating stable RNA products
Seven novel fibrillarin-associated snoRNAs, named U25–U31, are encoded within different introns of the unusually compact mammalian U22 host gene (UHG), which specifies the functional products of UHG.
U20, a novel small nucleolar RNA, is encoded in an intron of the nucleolin gene in mammals.
U20 RNA contains an extended region (21 nucleotides) of perfect complementarity with a phylogenetically conserved sequence in 18S rRNA, suggesting that U20 RNA may be involved in the formation of the small ribosomal subunit like nucleolin, the product of its host gene.
Processing of truncated mouse or human rRNA transcribed from ribosomal minigenes transfected into mouse cells.
It is shown that the major cis signals for pre-rRNA processing at the 18S rRNA/ITS 1 or the ITS2/28S r RNA junction involve solely a limited critical length of the respective mature rRNA and adjacent spacer sequences.
Novel intron-encoded small nucleolar RNAs with long sequence complementarities to mature rRNAs involved in ribosome biogenesis.
Phylogenetic evidences support the fundamental importance of the pairings of these three snoRNAs to pre-rRNA, which could be involved in a control of pre- rRNA folding during preribosome assembly.
Intron-encoded, antisense small nucleolar RNAs: the characterization of nine novel species points to their direct role as guides for the 2'-O-ribose methylation of rRNAs.
Nine additional members of the snoRNAs identified are intron-encoded, contain the characteristic box C and box D motifs and exhibit long complementarities to conserved sequences in mature rRNAs, supporting the conclusion that the duplex structure provides the 2'-O-methyltransferase with the appropriate site-specificity on the substrate.