{"doi":"10.1021/bi027297t","title":"Mechanistic Insights into Replication Across from Bulky DNA Adducts:  A Mutant Polymerase I Allows an <i>N</i>-Acetyl-2-aminofluorene Adduct To Be Accommodated during DNA Synthesis","abstract":null,"journal":"Biochemistry","year":2003,"id":642980,"datarank":0.41588830833596724,"base_score":2.772588722239781,"endowment":2.772588722239781,"self_citation_contribution":0.41588830833596724,"citation_network_contribution":0.0,"self_endowment_contribution":0.41588830833596724,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":15,"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":1672736,"name":"Louis J. Romano","orcid":null,"position":1,"is_corresponding":false},{"id":1672735,"name":"Samer Lone","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Mechanistic Insights into Replication Across from Bulky DNA Adducts:  A Mutant Polymerase I Allows an <i>N</i>-Acetyl-2-aminofluorene Adduct To Be Accommodated during DNA Synthesis","abstract":"The molecular mechanism that allows a polymerase to incorporate a nucleotide opposite a DNA lesion is not well-understood. One way to study this process is to characterize the altered molecular interactions that occur between the polymerase and a damaged template. Prior studies have determined the polymerase-template dissociation constants and used kinetic analyses and a protease digestion assay to measure the effect of various DNA adducts positioned in the active site of Klenow fragment (KF). Here, a mutator polymerase was used in which the tyrosine at position 766 of the KF has been replaced with a serine. This position is located at the junction of the fingers and palm domain and is thought to be involved in maintaining the active site geometry. The primer-template was modified with N-acetyl-2-aminofluorene (AAF), a well-studied carcinogenic adduct. The mutant polymerase displayed a significant increase in the rate of incorporation of the correct nucleotide opposite the adduct but was much less prone to incorporate an incorrect nucleotide relative to the wild-type polymerase. Both the wild-type and the mutant polymerase bound much more tightly to the AAF-modified primer-template; however, unlike the wild-type polymerase, the binding strength of the mutant was influenced by the presence of a dNTP. Moreover, the mutant polymerase was able to undergo a dNTP-induced conformational change when the AAF adduct was positioned in the active site, while the wild-type enzyme could not. A model is proposed in which the looser active site of the mutant is able to better accommodate the AAF adduct.","is_dataset_classified":null,"base_score":2.772588722239781,"endowment":2.772588722239781,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"12667073","pmcid":null,"openalex_id":"https://openalex.org/W1983319274","authors":[],"funders":[{"funder_name":"NCI NIH HHS","grant_id":"CA40605","title":null}],"total_grants":1,"fwci":0.4561,"citation_percentile":0.59839086,"influential_citations":0,"citation_trend":[{"year":2012,"count":1},{"year":2019,"count":1},{"year":2024,"count":1}],"oa_status":"closed","license":null,"oa_locations":[{"url":"https://pubs.acs.org/doi/pdf/10.1021/bi027297t","host_type":"publisher"},{"url":"https://doi.org/10.1021/bi027297t","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/12667073","host_type":"repository"}],"fields_of_study":["DNA Repair Mechanisms","DNA and Nucleic Acid Chemistry","Carcinogens and Genotoxicity Assessment"],"mesh_terms":["Binding Sites","Carcinogens","DNA Damage","DNA Polymerase I","DNA Replication","Escherichia coli","Fluorenes","Kinetics","Mutation","Protein Binding","Protein Conformation","Templates, Genetic","Trypsin","Tyrosine","2-Acetylaminofluorene","Mutagenesis, Site-Directed","DNA Primers","DNA Adducts"],"keywords":["Klenow fragment","DNA polymerase","Polymerase","DNA polymerase I","DNA clamp","DNA polymerase II","Primer (cosmetics)","Chemistry","Processivity","DNA polymerase beta","Molecular biology","Mutant","DNA replication","Active site","Biochemistry","DNA polymerase delta","DNA","Enzyme","Biology","Exonuclease","DNA repair","Polymerase chain reaction","Reverse transcriptase","Gene","Base excision repair"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-08T11:21:35.562097Z","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":[]}