Interaction of translation initiation factor eIF4G with eIF4A in the yeast Saccharomyces cerevisiae.
Eukaryotic initiation factor (eIF) 4A is an essential protein that, in conjunction with eIF4B, catalyzes the ATP-dependent melting of RNA secondary structure in the 5'-untranslated region of mRNA during translation initiation. In higher eukaryotes, eIF4A is assumed to be recruited to the mRNA through its interaction with eIF4G. However, the failure to detect this interaction in yeast brought into question the generality of this model. The work presented here demonstrates that yeast eIF4G interacts with eIF4A both in vivo and in vitro. The eIF4A-binding site was mapped to amino acids 542-883 of yeast eIF4G1. Expression in yeast cells of the eIF4G1 domain that binds eIF4A results in cell growth inhibition, and addition of this domain to an eIF4A-dependent in vitro system inhibits translation in a dose-dependent manner. Both in vitro translation and cell growth can be specifically restored by increasing the eIF4A concentration. These data demonstrate that yeast eIF4A and eIF4G interact and suggest that this interaction is required for translation and cell growth.
Dominguez
D
D
Institute for Biochemistry and Molecular Biology, University of Bern, Bühlstrasse 28, 3012 Bern, Switzerland. diana-ines.dominguez@mci.unibe.ch
Altmann
M
M
Benz
J
J
Baumann
U
U
Trachsel
H
H
D039601:Eukaryotic Initiation Factor-4A; D039603:Eukaryotic Initiation Factor-4G; D009690:Nucleic Acid Conformation; D010448:Peptide Initiation Factors / Q000378:metabolism*; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*
D039603:Eukaryotic Initiation Factor-4G; D010448:Peptide Initiation Factors; D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins; D039601:Eukaryotic Initiation Factor-4A