Rea1, a dynein-related nuclear AAA-ATPase, is involved in late rRNA processing and nuclear export of 60 S subunits.
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.
Galani
Kyriaki
K
Biochemie-Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld 328, 69120 Heidelberg, Germany.
Nissan
T Amar
TA
Petfalski
Elisabeth
E
Tollervey
David
D
Hurt
Ed
E
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry; D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000502:physiology*; D000483:Alleles; D015152:Blotting, Northern; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D021903:DNA, Ribosomal Spacer; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015636:Magnesium Chloride / Q000494:pharmacology; D008565:Membrane Proteins / Q000737:chemistry; D008954:Models, Biological; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry; D012335:RNA, Ribosomal / Q000378:metabolism*; D012340:RNA, Ribosomal, 5.8S / Q000737:chemistry; D011987:Receptors, Steroid; D012270:Ribosomes / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000502:physiology*; D012492:Salts / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D000098825:Oxysterol Binding Proteins
D018528:ATP-Binding Cassette Transporters; D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases; D021903:DNA, Ribosomal Spacer; D049452:Green Fluorescent Proteins; D015636:Magnesium Chloride; D008565:Membrane Proteins; D009841:Oligonucleotides; D012313:RNA; D012335:RNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; D011987:Receptors, Steroid; D029701:Saccharomyces cerevisiae Proteins; D012492:Salts; D012967:Sodium Dodecyl Sulfate; C087901:KES1 protein, S cerevisiae; D000098825:Oxysterol Binding Proteins; C098338:PXA1 protein, S cerevisiae; C496539:Rix1 protein, S cerevisiae; C496538:MDN1 protein, S cerevisiae