title: The h subunit of eIF3 promotes reinitiation competence during translation of mRNAs harboring upstream open reading frames.
abstract: Upstream open reading frames (uORFs) are protein coding elements in the 5' leader of messenger RNAs. uORFs generally inhibit translation of the main ORF because ribosomes that perform translation elongation suffer either permanent or conditional loss of reinitiation competence. After conditional loss, reinitiation competence may be regained by, at the minimum, reacquisition of a fresh methionyl-tRNA. The conserved h subunit of Arabidopsis eukaryotic initiation factor 3 (eIF3) mitigates the inhibitory effects of certain uORFs. Here, we define more precisely how this occurs, by combining gene expression data from mutated 5' leaders of Arabidopsis AtbZip11 (At4g34590) and yeast GCN4 with a computational model of translation initiation in wild-type and eif3h mutant plants. Of the four phylogenetically conserved uORFs in AtbZip11, three are inhibitory to translation, while one is anti-inhibitory. The mutation in eIF3h has no major effect on uORF start codon recognition. Instead, eIF3h supports efficient reinitiation after uORF translation. Modeling suggested that the permanent loss of reinitiation competence during uORF translation occurs at a faster rate in the mutant than in the wild type. Thus, eIF3h ensures that a fraction of uORF-translating ribosomes retain their competence to resume scanning. Experiments using the yeast GCN4 leader provided no evidence that eIF3h fosters tRNA reaquisition. Together, these results attribute a specific molecular function in translation initiation to an individual eIF3 subunit in a multicellular eukaryote.
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, The University of Tennessee, Knoxville, Tennessee 37996, USA.
authors.forename: Bijoyita
authors.initials: B
authors.lastname: Roy
authors.forename: Justin N
authors.initials: JN
authors.lastname: Vaughn
authors.forename: Byung-Hoon
authors.initials: BH
authors.lastname: Kim
authors.forename: Fujun
authors.initials: F
authors.lastname: Zhou
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gilchrist
authors.forename: Albrecht G
authors.initials: AG
authors.lastname: Von Arnim
mesh_terms: D020121:5' Untranslated Regions; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D018387:Codon, Initiator; D039621:Eukaryotic Initiation Factor-3 / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D016366:Open Reading Frames; D010442:Peptide Chain Initiation, Translational; D014176:Protein Biosynthesis; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; C112599:ATB2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D050976:Basic-Leucine Zipper Transcription Factors; D018387:Codon, Initiator; D039621:Eukaryotic Initiation Factor-3; C501391:GCN4 protein, S cerevisiae; D021122:Protein Subunits; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins