biostudies-literatureHoernes TPAustrian Science Fund FWF4865https://www.ebi.ac.uk/biostudies/studies/S-EPMC6242847biostudies-literatureUnknown9(1)The precise interplay between the mRNA codon and the tRNA anticodon is crucial for ensuring efficient and accurate translation by the ribosome. The insertion of RNA nucleobase derivatives in the mRNA allowed us to modulate the stability of the codon-anticodon interaction in the decoding site of bacterial and eukaryotic ribosomes, allowing an in-depth analysis of codon recognition. We found the hydrogen bond between the N¹ of purines and the N³ of pyrimidines to be sufficient for decoding of the first two codon nucleotides, whereas adequate stacking between the RNA bases is critical at the wobble position. Inosine, found in eukaryotic mRNAs, is an important example of destabilization of the codon-anticodon interaction. Whereas single inosines are efficiently translatedNature communicationsTranslation of non-standard codon nucleotides reveals minimal requirements for codon-anticodon interactions.PMC6242847P 28494P 22658-B12P 30486P 22658Faserl KHuttenhofer AJoseph SLindner HShi XErlacher MDMicura RWesthof EHobartner CJuen MASiewert AKremser JGasser CFuchs EHoernes TPKreutz CfalseTranslation of non-standard codon nucleotides reveals minimal requirements for codon-anticodon interactions.The precise interplay between the mRNA codon and the tRNA anticodon is crucial for ensuring efficient and accurate translation by the ribosome. The insertion of RNA nucleobase derivatives in the mRNA allowed us to modulate the stability of the codon-anticodon interaction in the decoding site of bacterial and eukaryotic ribosomes, allowing an in-depth analysis of codon recognition. We found the hydrogen bond between the N¹ of purines and the N³ of pyrimidines to be sufficient for decoding of the first two codon nucleotides, whereas adequate stacking between the RNA bases is critical at the wobble position. Inosine, found in eukaryotic mRNAs, is an important example of destabilization of the codon-anticodon interaction. Whereas single inosines are efficiently translated2018-01-01T00:00:00Z2018 Nov2026-05-06T04:02:19.805Z2019-03-27T00:08:33ZS-EPMC62428473045186110.1038/s41467-018-07321-8