Suppression of a temperature-sensitive cdc33 mutation of yeast by a multicopy plasmid expressing a Drosophila ribosomal protein.
The Saccharomyces cerevisiae cdc33ts4-2 mutant produces a temperature-sensitive allele of the cap-binding subunit of eukaryotic initiation factor-4F (also termed eIF-4E). From a Drosophila cDNA library constructed in a multicopy yeast shuttle vector, a clone was isolated which restored the ability to grow at elevated temperature to cdc33ts4-2 cells. The rescuing Drosophila clone encodes a small ribosomal subunit protein, which we name S15a based on its molecular weight and similarity with the Brassica napus S15a ribosomal protein. Transcription of the Drosophila gene, RpS15a, occurs at all developmental stages and is enhanced during oogenesis. The ribosomal protein gene is capable of suppressing other alleles of cdc33 but not an inactivation mutation, suggesting that suppression is dependent upon the presence of the temperature-sensitive eIF-4E protein. Supporting this, Western blot analysis shows that far more eIF-4E protein is present in cdc33 yeast cells expressing the RpS15a gene than lacking it. Levels of other unrelated proteins are unaffected. We propose therefore that the expression of high levels of the Drosophila S15a ribosomal protein in the cdc33 yeast cells leads to a selective stabilization of the temperature-sensitive eIF-4E protein, which accounts for the suppression phenomenon.
Lavoie
C
C
Department of Biology, McGill University, Montréal, Québec, Canada.
Tam
R
R
Clark
M
M
Lee
H
H
Sonenberg
N
N
Lasko
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D001937:Brassica / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004331:Drosophila melanogaster / Q000235:genetics*; D039561:Eukaryotic Initiation Factor-4E; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009154:Mutation; D010448:Peptide Initiation Factors / Q000378:metabolism; D010957:Plasmids; D012269:Ribosomal Proteins / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013489:Suppression, Genetic; D013696:Temperature; D014158:Transcription, Genetic
D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D012269:Ribosomal Proteins; C074488:ribosomal protein S15a; D004247:DNA