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Nucleic Acids Res ; 52(3): 1064-1079, 2024 Feb 09.
Article in English | MEDLINE | ID: mdl-38038264

ABSTRACT

mRNA translation is a fundamental process for life. Selection of the translation initiation site (TIS) is crucial, as it establishes the correct open reading frame for mRNA decoding. Studies in vertebrate mRNAs discovered that a purine at -3 and a G at +4 (where A of the AUG initiator codon is numbered + 1), promote TIS recognition. However, the TIS context in other eukaryotes has been poorly experimentally analyzed. We analyzed in vitro the influence of the -3, -2, -1 and + 4 positions of the TIS context in rabbit, Drosophila, wheat, and yeast. We observed that -3A conferred the best translational efficiency across these species. However, we found variability at the + 4 position for optimal translation. In addition, the Kozak motif that was defined from mammalian cells was only weakly predictive for wheat and essentially non-predictive for yeast. We discovered eight conserved sequences that significantly disfavored translation. Due to the big differences in translational efficiency observed among weak TIS context sequences, we define a novel category that we termed 'barren AUG context sequences (BACS)', which represent sequences disfavoring translation. Analysis of mRNA-ribosomal complexes structures provided insights into the function of BACS. The gene ontology of the BACS-containing mRNAs is presented.


Subject(s)
Codon, Initiator , Conserved Sequence , Protein Biosynthesis , Animals , Rabbits , Codon, Initiator/genetics , Mammals/genetics , Peptide Chain Initiation, Translational , RNA, Messenger/metabolism , Yeasts , Eukaryota/genetics , Eukaryota/metabolism
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