Utp23p is required for dissociation of snR30 small nucleolar RNP from preribosomal particles

Nucleic Acids Res. 2012 Apr;40(8):3641-52. doi: 10.1093/nar/gkr1213. Epub 2011 Dec 17.

Abstract

Yeast snR30 is an essential box H/ACA small nucleolar RNA (snoRNA) that promotes 18S rRNA processing through forming transient base-pairing interactions with the newly synthesized 35S pre-rRNA. By using a novel tandem RNA affinity selection approach, followed by coimmunoprecipitation and in vivo cross-linking experiments, we demonstrate that in addition to the four H/ACA core proteins, Cbf5p, Nhp2p, Nop10p and Gar1p, a fraction of snR30 specifically associates with the Utp23p and Kri1p nucleolar proteins. Depletion of Utp23p and Kri1p has no effect on the accumulation and recruitment of snR30 to the nascent pre-ribosomes. However, in the absence of Utp23p, the majority of snR30 accumulates in large pre-ribosomal particles. The retained snR30 is not base-paired with the 35S pre-rRNA, indicating that its aberrant tethering to nascent preribosomes is likely mediated by pre-ribosomal protein(s). Thus, Utp23p may promote conformational changes of the pre-ribosome, essential for snR30 release. Neither Utp23p nor Kri1p is required for recruitment of snR30 to the nascent pre-ribosome. On the contrary, depletion of snR30 prevents proper incorporation of both Utp23p and Kri1p into the 90S pre-ribosome containing the 35S pre-rRNA, indicating that snR30 plays a central role in the assembly of functionally active small subunit processome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / metabolism*
  • Nuclear Proteins / physiology
  • RNA Precursors / metabolism
  • RNA, Ribosomal / metabolism
  • RNA, Small Nucleolar / chemistry
  • RNA, Small Nucleolar / isolation & purification
  • RNA, Small Nucleolar / metabolism*
  • Ribonucleoproteins, Small Nucleolar / isolation & purification
  • Ribosomes / metabolism*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Saccharomyces cerevisiae Proteins / physiology

Substances

  • Nuclear Proteins
  • RNA Precursors
  • RNA, Ribosomal
  • RNA, Small Nucleolar
  • Ribonucleoproteins, Small Nucleolar
  • Saccharomyces cerevisiae Proteins
  • Utp23 protein, S cerevisiae