Box C/D snoRNP catalysed methylation is aided by additional pre-rRNA base-pairing

Robert Willem Van Nues, Sander Granneman, Grzegorz Kudla, Katherine Elizabeth Sloan, Matthew Chicken, David Tollervey, Nicholas James Watkins

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

2'-O-methylation of eukaryotic ribosomal RNA (r)RNA, essential for ribosome function, is catalysed by box C/D small nucleolar (sno)RNPs. The RNA components of these complexes (snoRNAs) contain one or two guide sequences, which, through base-pairing, select the rRNA modification site. Adjacent to the guide sequences are protein-binding sites (the C/D or C'/D' motifs). Analysis of >2000 yeast box C/D snoRNAs identified additional conserved sequences in many snoRNAs that are complementary to regions adjacent to the rRNA methylation site. This 'extra base-pairing' was also found in many human box C/D snoRNAs and can stimulate methylation by up to five-fold. Sequence analysis, combined with RNA-protein crosslinking in Saccharomyces cerevisiae, identified highly divergent box C'/D' motifs that are bound by snoRNP proteins. In vivo rRNA methylation assays showed these to be active. Our data suggest roles for non-catalytic subunits (Nop56 and Nop58) in rRNA binding and support an asymmetric model for box C/D snoRNP organization. The study provides novel insights into the extent of the snoRNA-rRNA interactions required for efficient methylation and the structural organization of the snoRNPs.
Original languageEnglish
Pages (from-to)2420-2430
Number of pages11
JournalEMBO Journal
Volume30
Issue number12
DOIs
Publication statusPublished - 15 Jun 2011

Keywords / Materials (for Non-textual outputs)

  • methylation
  • ribosome
  • rRNA
  • snoRNA
  • snoRNP

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