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
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP-Binding Cassette Transporters / chemistry
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ATPases Associated with Diverse Cellular Activities
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Adenosine Triphosphatases / chemistry*
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Adenosine Triphosphatases / physiology*
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Alleles
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Blotting, Northern
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Blotting, Western
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Cell Nucleus / metabolism
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Cytoplasm / metabolism
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DNA, Ribosomal Spacer
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Genes, Reporter
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Green Fluorescent Proteins / metabolism
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Magnesium Chloride / pharmacology
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Membrane Proteins / chemistry
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Models, Biological
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Mutation
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Oligonucleotides / chemistry
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Plasmids / metabolism
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Protein Structure, Tertiary
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RNA / chemistry
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RNA, Ribosomal / metabolism*
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RNA, Ribosomal, 5.8S / chemistry
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Receptors, Steroid
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Ribosomes / chemistry*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / physiology*
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Salts / pharmacology
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Sodium Dodecyl Sulfate / chemistry
Substances
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ATP-Binding Cassette Transporters
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DNA, Ribosomal Spacer
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KES1 protein, S cerevisiae
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Membrane Proteins
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Oligonucleotides
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PXA1 protein, S cerevisiae
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RNA, Ribosomal
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RNA, Ribosomal, 5.8S
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Receptors, Steroid
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Rix1 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Salts
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Magnesium Chloride
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Green Fluorescent Proteins
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Sodium Dodecyl Sulfate
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RNA
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Adenosine Triphosphatases
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MDN1 protein, S cerevisiae
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ATPases Associated with Diverse Cellular Activities