{"doi":"10.1101/2022.06.13.495988","title":"N1-Methylpseudouridine and pseudouridine modifications modulate mRNA decoding during translation","abstract":"The ribosome relies on hydrogen bonding interactions between mRNA codons and incoming aminoacyl-tRNAs to ensure rapid and accurate protein production. The inclusion of chemically modified bases into mRNAs has the potential to alter the strength and pattern of hydrogen bonding between mRNAs and aminoacyl-tRNAs to alter protein synthesis. We investigated how the Nl-methylpseudouridine (m 1 Ψ) modification, commonly incorporated into therapeutic and vaccine mRNA sequences, influences the ability of codons to react with cognate and near-cognate tRNAs and release factors. We find that the presence of a single m 1 Ψ does not substantially change the rate constants for amino acid addition by cognate tRNAs or termination by release factors. However, insertion of m 1 Ψ can affect the selection of near-cognate tRNAs both in vitro and in human cells. Our observations demonstrate that m 1 Ψ, and the related naturally occurring pseudouridine (Ψ) modification, exhibit the ability to both increase and decrease the extent of amino acid misincorporation in a codon-position and tRNA dependent manner. To ascertain the chemical logic for our biochemical and cellular observations, we computationally modeled tRNA Ile(GAU) bound to unmodified and m 1 Ψ- or Ψ-modified phenylalanine codons (UUU). Our modeling suggests that changes in the energetics of mRNA:tRNA interactions largely correlate with the context specificity of Ile-miscoding events we observe on Ψ and m 1 Ψ containing Phe codons. These studies reveal that the sequence context of a given modification within an mRNA plays a large role in determining how (and if) the modification impacts the number and distribution of proteoforms synthesized by the ribosome.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":297966,"datarank":0.31191623125197543,"base_score":2.0794415416798357,"endowment":2.0794415416798357,"self_citation_contribution":0.31191623125197543,"citation_network_contribution":0.0,"self_endowment_contribution":0.31191623125197543,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":7,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.96,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":987010,"name":"Lili Mitchell","orcid":null,"position":1,"is_corresponding":false},{"id":833110,"name":"Indrajit Deb","orcid":"0000-0002-2722-582X","position":2,"is_corresponding":false},{"id":357088,"name":"Bijoyita Roy","orcid":"0000-0003-1061-5482","position":3,"is_corresponding":false},{"id":600007,"name":"Aaron T. Frank","orcid":"0000-0002-7782-2200","position":4,"is_corresponding":false},{"id":304098,"name":"Kristin S. Koutmou","orcid":"0000-0002-7763-9262","position":5,"is_corresponding":false},{"id":304097,"name":"Jeremy Monroe","orcid":"0000-0002-1901-8345","position":0,"is_corresponding":true}],"reference_count":93,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T00:31:31.270564Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}