Functional requirement for symmetric assembly of archaeal box C/D small ribonucleoprotein particles

Rumana Rashid, Mohamed Aittaleb, Qiong Chen, Katharina Spiegel, Borries Demeler, Hong Li

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

Box C/D small ribonucleoprotein particles (sRNPs) are archaeal homologs of small nucleolar ribonucleoprotein particles (snoRNPs) in eukaryotes that are responsible for site specific 2′-O-methylation of ribosomal and transfer RNAs. The function of box C/D sRNPs is characterized by step-wise assembly of three core proteins around a box C/D RNA that include fibrillarin, Nop5p, and L7Ae. The most distinct structural feature in all box C/D RNAs is the presence of two conserved box C/D motifs accompanied by often a single, and sometimes two, antisense elements located immediately upstream of either the D or D′ box. Despite this asymmetric distribution of antisense elements, the bipartite feature of the box C/D motifs appears to be in pleasing agreement with a recently reported three-dimensional structure of the core protein complex between fibrillarin and Nop5p. This investigates functional implications of the symmetric features both in box C/D RNAs and in the fibrillarin-Nop5p complex. Site-directed mutagenesis was employed to generate box C/D RNAs lacking one of the two box C/D motifs and a mutant fibrillarin-Nop5p complex deficient in self-association. The ability of the mutated components to assemble and to direct methyl transfer reactions was assessed by gel mobility-shift, analytical ultracentrifugation, and in vitro catalysis studies. The results presented here suggest that, while a box C/D sRNP is capable of asymmetrical assembly, the symmetries in both the box C/D RNA and in the fibrillarin-Nop5p complex are required for efficient catalysis. These findings underscore the importance of functional assembly in methyl transfer reactions.

Original languageEnglish (US)
Pages (from-to)295-306
Number of pages12
JournalJournal of Molecular Biology
Volume333
Issue number2
DOIs
StatePublished - Oct 17 2003

Keywords

  • 2′-O-methylation
  • RNA methyltransferase
  • RNP assembly
  • Small guide RNA
  • rRNA biogenesis

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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