A region rich in aspartic acid, arginine, tyrosine, and glycine (DRYG) mediates eukaryotic initiation factor 4B (eIF4B) self-association and interaction with eIF3.
The binding of mRNA to the ribosome is mediated by eukaryotic initiation factors eukaryotic initiation factor 4F (eIF4F), eIF4B, eIF4A, and eIF3, eIF4F binds to the mRNA cap structure and, in combination with eIF4B, is believed to unwind the secondary structure in the 5' untranslated region to facilitate ribosome binding. eIF3 associates with the 40S ribosomal subunit prior to mRNA binding. eIF4B copurifies with eIF3 and eIF4F through several purification steps, suggesting the involvement of a multisubunit complex during translation initiation. To understand the mechanism by which eIF4B promotes 40S ribosome binding to the mRNA, we studied its interactions with partner proteins by using a filter overlay (protein-protein [far Western]) assay and the two-hybrid system. In this report, we show that eIF4B self-associates and also interacts directly with the p170 subunit of eIF3. A region rich in aspartic acid, arginine, tyrosine, and glycine, termed the DRYG domain, is sufficient for self-association of eIF4B, both in vitro and in vivo, and for interaction with the p170 subunit of eIF3. These experiments suggest that eIF4B participates in mRNA-ribosome binding by acting as an intermediary between the mRNA and eIF3, via a direct interaction with the p170 subunit of eIF3.
Méthot
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Department of Biochemistry, McGill University, Montréal, Québec, Canada.
Song
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MS
Sonenberg
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D000818:Animals; D001120:Arginine; D001224:Aspartic Acid; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D039621:Eukaryotic Initiation Factor-3; D039642:Eukaryotic Initiation Factors; D005998:Glycine; D006801:Humans; D046911:Macromolecular Substances; D008961:Models, Structural; D010448:Peptide Initiation Factors / Q000737:chemistry* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D014443:Tyrosine
D017931:DNA Primers; D004268:DNA-Binding Proteins; D039621:Eukaryotic Initiation Factor-3; D039642:Eukaryotic Initiation Factors; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins; C031799:eIF-4B; D001224:Aspartic Acid; D014443:Tyrosine; D001120:Arginine; D005998:Glycine