Rea1, a dynein-related nuclear AAA-ATPase, is involved in late rRNA processing and nuclear export of 60 S subunits

J Biol Chem. 2004 Dec 31;279(53):55411-8. doi: 10.1074/jbc.M406876200. Epub 2004 Nov 3.

Abstract

Rea1, the largest predicted protein in the yeast genome, is a member of the AAA(+) family of ATPases and is associated with pre-60 S ribosomes. Here we report that Rea1 is required for maturation and nuclear export of the pre-60 S subunit. Rea1 exhibits a predominantly nucleoplasmic localization and is present in a late pre-60 S particle together with members of the Rix1 complex. To study the role of Rea1 in ribosome biogenesis, we generated a repressible GAL::REA1 strain and temperature-sensitive rea1 alleles. In vivo depletion of Rea1 results in the significant reduction of mature 60 S subunits concomitant with defects in pre-rRNA processing and late pre-60 S ribosome stability following ITS2 cleavage and prior to the generation of mature 5.8 S rRNA. Strains depleted of the components of the Rix1 complex (Rix1, Ipi1, and Ipi3) showed similar defects. Using an in vivo 60 S subunit export assay, a strong accumulation of the large subunit reporter Rpl25-GFP (green fluorescent protein) in the nucleus and at the nuclear periphery was seen in rea1 mutants at restrictive conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry
  • ATPases Associated with Diverse Cellular Activities
  • Adenosine Triphosphatases / chemistry*
  • Adenosine Triphosphatases / physiology*
  • Alleles
  • Blotting, Northern
  • Blotting, Western
  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • DNA, Ribosomal Spacer
  • Genes, Reporter
  • Green Fluorescent Proteins / metabolism
  • Magnesium Chloride / pharmacology
  • Membrane Proteins / chemistry
  • Models, Biological
  • Mutation
  • Oligonucleotides / chemistry
  • Plasmids / metabolism
  • Protein Structure, Tertiary
  • RNA / chemistry
  • RNA, Ribosomal / metabolism*
  • RNA, Ribosomal, 5.8S / chemistry
  • Receptors, Steroid
  • Ribosomes / chemistry*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / physiology*
  • Salts / pharmacology
  • Sodium Dodecyl Sulfate / chemistry

Substances

  • ATP-Binding Cassette Transporters
  • DNA, Ribosomal Spacer
  • KES1 protein, S cerevisiae
  • Membrane Proteins
  • Oligonucleotides
  • PXA1 protein, S cerevisiae
  • RNA, Ribosomal
  • RNA, Ribosomal, 5.8S
  • Receptors, Steroid
  • Rix1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Salts
  • Magnesium Chloride
  • Green Fluorescent Proteins
  • Sodium Dodecyl Sulfate
  • RNA
  • Adenosine Triphosphatases
  • MDN1 protein, S cerevisiae
  • ATPases Associated with Diverse Cellular Activities