{"doi":"10.1016/s0076-6879(07)30007-4","title":"Assembly and Analysis of Eukaryotic Translation Initiation Complexes","abstract":null,"journal":"Methods in Enzymology","year":2007,"id":682521,"datarank":0.7412463633913957,"base_score":4.941642422609304,"endowment":4.941642422609304,"self_citation_contribution":0.7412463633913957,"citation_network_contribution":0.0,"self_endowment_contribution":0.7412463633913957,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":139,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"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":137803,"name":"Anett Unbehaun","orcid":null,"position":1,"is_corresponding":false},{"id":136910,"name":"Christopher U.T. Hellen","orcid":null,"position":2,"is_corresponding":false},{"id":136906,"name":"Tatyana V. Pestova","orcid":null,"position":3,"is_corresponding":false},{"id":1437643,"name":"Andrey V. Pisarev","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Assembly and Analysis of Eukaryotic Translation Initiation Complexes","abstract":"The canonical initiation process is the most complex aspect of translation in eukaryotes. It involves the coordinated interactions of at least 11 eukaryotic initiation factors, 40S and 60S ribosomal subunits, mRNA, and aminoacylated initiator tRNA (Met-tRNA(i)(Met)), as well as binding and hydrolysis of GTP and ATP. The factor requirements for many individual steps in this process, including scanning, initiation codon recognition, and ribosomal subunit joining, have until recently been obscure. We established the factor requirements for these steps by reconstituting the initiation process in vitro from individual purified components of the translation apparatus and developed approaches to explain the mechanism of individual steps and the roles of individual factors and to characterize the structure of initiation complexes. Here we describe protocols for the purification of native initiation factors and for expression and purification of active recombinant forms of all single subunit initiation factors, for the reconstitution of the initiation process, and for determination of the position of ribosomal complexes on mRNA by primer extension inhibition (\"toe printing\"). We also describe protocols for site-directed ultraviolet (UV) cross-linking to determine the interactions of individual nucleotides in mRNA with components of the initiation complex and for directed hydroxyl radical probing to determine the position of initiation factors on the ribosome.","is_dataset_classified":null,"base_score":4.941642422609304,"endowment":4.941642422609304,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"17913638","pmcid":null,"openalex_id":"https://openalex.org/W1525224242","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM59660","title":null}],"total_grants":1,"fwci":11.2501,"citation_percentile":0.98461538,"influential_citations":9,"citation_trend":[{"year":2012,"count":8},{"year":2013,"count":9},{"year":2014,"count":6},{"year":2015,"count":6},{"year":2016,"count":5},{"year":2017,"count":7},{"year":2018,"count":11},{"year":2019,"count":10},{"year":2020,"count":8},{"year":2021,"count":10},{"year":2022,"count":4},{"year":2023,"count":5},{"year":2024,"count":9},{"year":2025,"count":6},{"year":2026,"count":6}],"oa_status":"closed","license":null,"oa_locations":[{"url":"https://doi.org/10.1016/s0076-6879(07)30007-4","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/17913638","host_type":"repository"}],"fields_of_study":["RNA and protein synthesis mechanisms","RNA modifications and cancer","RNA Research and Splicing","Biology","Medicine","Animals","Cross-Linking Reagents","Eukaryotic Initiation Factors","Globins","Hydroxyl Radical","Macromolecular Substances","Methionine-tRNA Ligase","Protein Biosynthesis","Protein Isoforms","RNA, Messenger","RNA, Ribosomal","RNA, Transfer","Rabbits","Recombinant Proteins","Ribosome Subunits, Large, Eukaryotic","Ribosome Subunits, Small, Eukaryotic"],"mesh_terms":["Animals","Cross-Linking Reagents","Globins","Methionine-tRNA Ligase","Rabbits","Recombinant Proteins","RNA, Messenger","RNA, Ribosomal","RNA, Transfer","Protein Biosynthesis","Hydroxyl Radical","Protein Isoforms","Eukaryotic Initiation Factors","Macromolecular Substances","Ribosome Subunits, Small, Eukaryotic","Ribosome Subunits, Large, Eukaryotic"],"keywords":["Initiation factor","Eukaryotic initiation factor","Eukaryotic translation","eIF2","Translation (biology)","Eukaryotic Ribosome","Eukaryotic Small Ribosomal Subunit","Ribosome","Biology","Transfer RNA","Ribosomal RNA","Internal ribosome entry site","Biochemistry","Cell biology","Messenger RNA","RNA","Gene"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-17T20:39:32.269699Z","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":[]}