Interactions of the eIF-4F subunits in the yeast Saccharomyces cerevisiae.
Recognition of the cap structure at the 5' end of mRNA is one of the first events in initiation of eukaryotic translation. This step is mediated by the translation initiation factor 4F (eIF-4F). In mammalian cells this factor is composed of the cap-binding protein eIF-4E, eIF-4A, and a 220-kDa polypeptide. In yeast Saccharomyces cerevisiae, eIF-4E is found associated with a 150-kDa protein (p150) and a 20-kDa protein (p20). The resulting protein complex is proposed to represent yeast eIF-4F. To study the functions of p150 and p20 and their interaction with eIF-4E, we disrupted the genes encoding p150 and p20 and analyzed the effects on protein complex formation and cell viability. Yeast cells with single and double disruptions of the genes encoding p150 and p20 are viable, but p150 single and p150/p20 double disruptions show a slow growth phenotype. Gel chromatography and immunoadsorption experiments with a monoclonal anti-eIF-4E antibody coupled to protein G-Sepharose show that both p150 and p20 bind independently of each other to eIF-4E.
Lanker
S
S
Institut für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
Müller
P P
PP
Altmann
M
M
Goyer
C
C
Sonenberg
N
N
Trachsel
H
H
D000911:Antibodies, Monoclonal; D015139:Blotting, Southern; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D039562:Eukaryotic Initiation Factor-4F; D005800:Genes, Fungal; D005838:Genotype; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004271:DNA, Fungal; D039562:Eukaryotic Initiation Factor-4F; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors