{"doi":"10.1101/2023.09.29.560218","title":"Human eukaryotic initiation factor 4G directly binds the 40S ribosomal subunit to promote efficient translation","abstract":"<jats:title>ABSTRACT</jats:title>\n                <jats:p>\n                  Messenger RNA (mRNA) recruitment to the 40S ribosomal subunit is mediated by eukaryotic initiation factor 4F (eIF4F). This complex includes 3 subunits: eIF4E (m\n                  <jats:sup>7</jats:sup>\n                  G cap binding protein), eIF4A (DEAD box helicase), and eIF4G. Mammalian eIF4G is a scaffold that coordinates the activities of eIF4E and eIF4A and provides a bridge to connect the mRNA and 40S ribosomal subunit through its interaction with eIF3. While the roles of many eIF4G binding domains are relatively clear, the precise function of RNA binding by eIF4G remains to be elucidated. In this work, we used an eIF4G-dependent translation assay to reveal that the RNA binding domain (eIF4G-RBD; amino acids 682–720) stimulates translation. This stimulating activity is observed when eIF4G is independently tethered to an internal region of the mRNA, suggesting that the eIF4G-RBD promotes translation by a mechanism that is independent of the m\n                  <jats:sup>7</jats:sup>\n                  G cap and mRNA tethering. Using a kinetic helicase assay, we show that the eIF4G-RBD has a minimal effect on eIF4A helicase activity, demonstrating that the eIF4G-RBD is not required to coordinate eIF4F-dependent duplex unwinding. Unexpectedly, native gel electrophoresis and fluorescence polarization assays reveal a previously unidentified direct interaction between eIF4G and the 40S subunit. Using binding assays, our data show that this 40S subunit interaction is separate from the previously characterized interaction between eIF4G and eIF3. Thus, our work reveals how eIF4F can bind to the 40S subunit using eIF3-dependent and eIF3-independent binding domains to promote translation initiation.\n                </jats:p>","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":null,"id":22375,"datarank":0.2634209100495159,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.022005223184400836,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.022005223184400836,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":4,"citers_with_citation_signal":2,"citers_with_endowment":2,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":138408,"name":"Christopher S. Fraser","orcid":null,"position":1,"is_corresponding":false},{"id":138403,"name":"Nancy Villa","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"37808713","pmcid":null,"openalex_id":"https://openalex.org/W4387234050","authors":[],"funders":[{"funder_name":"National Institutes of Health","grant_id":"5R35GM152137-02","title":"The mechanisms of mRNA recruitment to the human ribosome"},{"funder_name":"National Institutes of Health","grant_id":"5R01GM092927-05","title":"The mechanism of mRNA recruitment to the human ribosome."}],"total_grants":2,"fwci":null,"citation_percentile":null,"influential_citations":1,"citation_trend":[{"year":2024,"count":3},{"year":2025,"count":1}],"oa_status":"green","license":"CC BY","oa_locations":[{"url":"https://www.biorxiv.org/content/biorxiv/early/2023/09/30/2023.09.29.560218.full.pdf","host_type":"repository"},{"url":"http://www.jbc.org/article/S0021925824017393/pdf","host_type":"GREEN"},{"url":"https://www.biorxiv.org/content/biorxiv/early/2023/09/30/2023.09.29.560218.full.pdf","host_type":"repository"},{"url":"https://syndication.highwire.org/content/doi/10.1101/2023.09.29.560218","host_type":"publisher"},{"url":"https://doi.org/10.1101/2023.09.29.560218","host_type":"repository"},{"url":"https://pubmed.ncbi.nlm.nih.gov/37808713","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/10557762","host_type":"repository"},{"url":"https://doi.org/10.1016/j.jbc.2024.107242","host_type":""},{"url":"https://pubmed.ncbi.nlm.nih.gov/38569933","host_type":""},{"url":"http://dx.doi.org/10.1016/j.jbc.2024.107242","host_type":""},{"url":"https://escholarship.org/uc/item/6f41x4dj","host_type":""},{"url":"https://escholarship.org/content/qt6f41x4dj/qt6f41x4dj.pdf","host_type":""},{"url":"https://doi.org/https://doi.org/10.1016/j.jbc.2024.107242","host_type":""}],"fields_of_study":["RNA and protein synthesis mechanisms","RNA Research and Splicing","RNA modifications and cancer","Biology","Medicine"],"mesh_terms":[],"keywords":["EIF4G","Eukaryotic Small Ribosomal Subunit","EIF4E","eIF4A","Initiation factor","Biology","Eukaryotic initiation factor","Eukaryotic translation","RNA Helicase A","Cell biology","Protein subunit","Messenger RNA","Molecular biology","RNA","Helicase","Translation (biology)","Biochemistry","Gene","570","Biochemistry & Molecular Biology","Biomedical and clinical sciences","mRNA","Eukaryotic Initiation Factor-3","Messenger","Small","RNA-binding","translation initiation","Medical and Health Sciences","Protein Domains","616","Genetics","Ribosome Subunits","Humans","RNA, Messenger","Ribosome Subunits, Small, Eukaryotic","Biological Sciences","Eukaryotic Initiation Factor-4E","Chemical sciences","eIF4F","Protein Biosynthesis","Eukaryotic","Biochemistry and Cell Biology","Generic health relevance","Eukaryotic Initiation Factor-4G","Research Article","Protein Binding"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-06T21:43:55.847664Z","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":[]}