Fibrillarin and Nop56 interact before being co-assembled in box C/D snoRNPs

Exp Cell Res. 2009 Apr 1;315(6):928-42. doi: 10.1016/j.yexcr.2009.01.016. Epub 2009 Jan 30.

Abstract

Small nucleolar RNAs play crucial roles in ribosome biogenesis. They guide folding, site-specific nucleotide modifications and participate in cleavage of precursor ribosomal RNAs. To better understand how the biogenesis of the box C/D small nucleolar RNPs (snoRNPs) occur in a cellular context, we used a new approach based on the possibility of relocalizing a given nuclear complex by adding an affinity tag for B23 to one component of this complex. We selectively delocalized each core box C/D protein, namely 15.5kD, Nop56, Nop58 and fibrillarin, and analyzed the effect of such changes on other components of the box C/D snoRNPs. We show that modifying the localization and the mobility of core box C/D proteins impairs their association with box C/D snoRNPs. In addition, we demonstrate that fibrillarin and Nop56 directly interact in vivo. This interaction, indispensable for the association of both proteins with the box C/D snoRNPs, does not involve the glycine- and arginine-rich domain or the RNA-binding domain but the alpha-helix domain of fibrillarin. In addition, no RNA seems required to maintain fibrillarin-Nop56 interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Fluorescence Recovery After Photobleaching
  • HeLa Cells
  • Humans
  • In Situ Hybridization, Fluorescence
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • RNA, Small Nucleolar / genetics
  • RNA, Small Nucleolar / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • NOLC1 protein, human
  • NOP56 protein, human
  • Nuclear Proteins
  • Phosphoproteins
  • RNA, Small Interfering
  • RNA, Small Nucleolar
  • Recombinant Fusion Proteins
  • fibrillarin