title: eIF1 controls multiple steps in start codon recognition during eukaryotic translation initiation.
abstract: Eukaryotic translation initiation factor (eIF) 1 is a central mediator of start codon recognition. Dissociation of eIF1 from the preinitiation complex (PIC) allows release of phosphate from the G-protein factor eIF2, triggering downstream events in initiation. Mutations that weaken binding of eIF1 to the PIC decrease the fidelity of start codon recognition (Sui(-) phenotype) by allowing increased eIF1 release at non-AUG codons. Consistent with this, overexpression of these mutant proteins suppresses their Sui(-) phenotypes. Here, we have examined mutations at the penultimate residue of eIF1, G107, that produce Sui(-) phenotypes without increasing the rate of eIF1 release. We provide evidence that, in addition to its role in gating phosphate release, dissociation of eIF1 triggers conversion from an open, scanning-competent state of the PIC to a stable, closed one. We also show that eIF5 antagonizes binding of eIF1 to the complex and that key interactions of eIF1 with its partners are modulated by the charge at and around G107. Our data indicate that eIF1 plays multiple roles in start codon recognition and suggest that prior to AUG recognition it prevents eIF5 from binding to a key site in the PIC required for triggering downstream events.
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Jagpreet S
authors.initials: JS
authors.lastname: Nanda
authors.forename: Yuen-Nei
authors.initials: YN
authors.lastname: Cheung
authors.forename: Julie E
authors.initials: JE
authors.lastname: Takacs
authors.forename: Pilar
authors.initials: P
authors.lastname: Martin-Marcos
authors.forename: Adesh K
authors.initials: AK
authors.lastname: Saini
authors.forename: Alan G
authors.initials: AG
authors.lastname: Hinnebusch
authors.forename: Jon R
authors.initials: JR
authors.lastname: Lorsch
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D018387:Codon, Initiator / Q000235:genetics / Q000378:metabolism*; D015977:Eukaryotic Initiation Factor-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010442:Peptide Chain Initiation, Translational; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D018387:Codon, Initiator; D015977:Eukaryotic Initiation Factor-1; C026022:eukaryotic peptide initiation factor-1A