Rpg1, the Saccharomyces cerevisiae homologue of the largest subunit of mammalian translation initiation factor 3, is required for translational activity.
Eukaryotic initiation factor 3 (eIF3) consists of at least eight subunits and plays a key role in the formation of the 43 S preinitiation complex by dissociating 40 and 60 S ribosomal subunits, stabilizing the ternary complex, and promoting mRNA binding to 40 S ribosomal subunits. The product of the Saccharomyces cerevisiae RPG1 gene has been described as encoding a protein required for passage through the G1 phase of the cell cycle and exhibiting significant sequence similarity to the largest subunit of human eIF3. Here we show that under nondenaturing conditions, Rpg1p copurifies with a known yeast eIF3 subunit, Prt1p. An anti-Rpg1p antibody co-immunoprecipitates Prt1p, and an antibody directed against the Myc tag of a tagged version of Prt1p co-immunoprecipitates Rpg1p, demonstrating that both proteins are present in the same complex. A cell-free translation system derived from the temperature-sensitive rpg1-1 mutant strain becomes inactivated by incubation at 37 degreesC, and its activity can be restored by the addition of the Rpg1-containing protein complex. Finally, the rpg1-1 temperature-sensitive mutant strain shows a dramatic reduction of the polysome/monosome ratio upon shift to the restrictive temperature. These data show that Rpg1p is an authentic eIF3 subunit and plays an important role in the initiation step of translation.
Valásek
L
L
Vienna Biocenter, Institute of Biochemistry and Molecular Cell Biology, University of Vienna, A-1030 Vienna, Austria.
Trachsel
H
H
Hasek
J
J
Ruis
H
H
D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D039621:Eukaryotic Initiation Factor-3; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D010448:Peptide Initiation Factors / Q000235:genetics*; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins
D018797:Cell Cycle Proteins; D039621:Eukaryotic Initiation Factor-3; D005656:Fungal Proteins; D010448:Peptide Initiation Factors; C050998:Prt1 protein, S cerevisiae; C111889:RPG1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins