{"doi":"10.1093/nar/gkab416","title":"Budding yeast Rap1, but not telomeric DNA, is inhibitory for multiple stages of DNA replication in vitro","abstract":"<jats:title>Abstract</jats:title>\n               <jats:p>Telomeres are copied and reassembled each cell division cycle through a multistep process called telomere replication. Most telomeric DNA is duplicated semiconservatively during this process, but replication forks frequently pause or stall at telomeres in yeast, mouse and human cells, potentially causing chronic telomere shortening or loss in a single cell cycle. We have investigated the cause of this effect by examining the replication of telomeric templates in vitro. Using a reconstituted assay for eukaryotic DNA replication in which a complete eukaryotic replisome is assembled and activated with purified proteins, we show that budding yeast telomeric DNA is efficiently duplicated in vitro unless the telomere binding protein Rap1 is present. Rap1 acts as a roadblock that prevents replisome progression and leading strand synthesis, but also potently inhibits lagging strand telomere replication behind the fork. Both defects can be mitigated by the Pif1 helicase. Our results suggest that GC-rich sequences do not inhibit DNA replication per se, and that in the absence of accessory factors, telomere binding proteins can inhibit multiple, distinct steps in the replication process.</jats:p>","journal":"Nucleic Acids Research","year":2021,"id":653957,"datarank":0.9779370182179077,"base_score":3.332204510175204,"endowment":3.332204510175204,"self_citation_contribution":0.49983067652628066,"citation_network_contribution":0.4781063416916271,"self_endowment_contribution":0.49983067652628066,"citer_contribution":0.4781063416916271,"corpus_percentile":null,"corpus_rank":null,"citation_count":27,"citer_count":23,"citers_with_citation_signal":21,"citers_with_endowment":21,"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":609024,"name":"John F.X. Diffley","orcid":"0000-0001-5184-7680","position":1,"is_corresponding":false},{"id":1232623,"name":"Max E. Douglas","orcid":"0000-0003-1186-7163","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Budding yeast Rap1, but not telomeric DNA, is inhibitory for multiple stages of DNA replication in vitro","abstract":"<jats:title>Abstract</jats:title>\n               <jats:p>Telomeres are copied and reassembled each cell division cycle through a multistep process called telomere replication. Most telomeric DNA is duplicated semiconservatively during this process, but replication forks frequently pause or stall at telomeres in yeast, mouse and human cells, potentially causing chronic telomere shortening or loss in a single cell cycle. We have investigated the cause of this effect by examining the replication of telomeric templates in vitro. Using a reconstituted assay for eukaryotic DNA replication in which a complete eukaryotic replisome is assembled and activated with purified proteins, we show that budding yeast telomeric DNA is efficiently duplicated in vitro unless the telomere binding protein Rap1 is present. Rap1 acts as a roadblock that prevents replisome progression and leading strand synthesis, but also potently inhibits lagging strand telomere replication behind the fork. Both defects can be mitigated by the Pif1 helicase. Our results suggest that GC-rich sequences do not inhibit DNA replication per se, and that in the absence of accessory factors, telomere binding proteins can inhibit multiple, distinct steps in the replication process.</jats:p>","is_dataset_classified":null,"base_score":3.295836866004329,"endowment":3.295836866004329,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"34048583","pmcid":"PMC8191780","openalex_id":"https://openalex.org/W3164612803","authors":[],"funders":[{"funder_name":"Cancer Research UK","grant_id":"C68409/A28129","title":null},{"funder_name":"Cancer Research UK","grant_id":"FC001065","title":null},{"funder_name":"Cancer Research UK","grant_id":"FC001066","title":null},{"funder_name":"Wellcome Trust","grant_id":"106252/Z/14/Z","title":null},{"funder_name":"European Research Council","grant_id":"669424-CHROMOREP","title":null},{"funder_name":"The Francis Crick Institute","grant_id":"10066","title":null},{"funder_name":"European Research Council","grant_id":"669424","title":"Reconstitution of Chromosome Replication and Epigenetic Inheritance"},{"funder_name":"Cancer Research UK","grant_id":"28129","title":null},{"funder_name":"The Francis Crick Institute","grant_id":"10668","title":null},{"funder_name":"Wellcome Trust","grant_id":"106252","title":"How the eukaryotic replicative DNA helicase is loaded and activated."},{"funder_name":"Francis Crick Institute","grant_id":"","title":null}],"total_grants":11,"fwci":1.9411,"citation_percentile":0.86247996,"influential_citations":0,"citation_trend":[{"year":2020,"count":1},{"year":2022,"count":7},{"year":2023,"count":3},{"year":2024,"count":7},{"year":2025,"count":6},{"year":2026,"count":2}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://academic.oup.com/nar/article-pdf/49/10/5671/38592670/gkab416.pdf","host_type":"journal"},{"url":"https://academic.oup.com/nar/article-pdf/49/10/5671/38592670/gkab416.pdf","host_type":"publisher"},{"url":"http://academic.oup.com/nar/article-pdf/49/10/5671/38592670/gkab416.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1093/nar/gkab416","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/34048583","host_type":"repository"},{"url":"https://figshare.com/articles/journal_contribution/Budding_yeast_Rap1_but_not_telomeric_DNA_is_inhibitory_for_multiple_stages_of_DNA_replication_in_vitro_/14790669","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/8191780","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC8191780","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC8191780?pdf=render","host_type":"Europe_PMC"},{"url":"http://dx.doi.org/10.1093/nar/gkab416","host_type":""},{"url":"https://dx.doi.org/10.1093/nar/gkab416","host_type":""},{"url":"https://doi.org/https://doi.org/10.1093/nar/gkab416","host_type":""}],"fields_of_study":["Telomeres, Telomerase, and Senescence","Genetics, Aging, and Longevity in Model Organisms","DNA Repair Mechanisms","0301 basic medicine","0303 health sciences","03 medical and health sciences","Base Composition","DNA Helicases","DNA Replication","Gene Expression","In Vitro Techniques","Recombinant Proteins","Saccharomyces cerevisiae Proteins","Saccharomycetales","Shelterin Complex","Telomere","Telomere-Binding Proteins","Transcription Factors"],"mesh_terms":["Shelterin Complex","Base Composition","DNA Replication","DNA Helicases","Saccharomycetales","Recombinant Proteins","Transcription Factors","Gene Expression","Telomere","Saccharomyces cerevisiae Proteins","Telomere-Binding Proteins","In Vitro Techniques"],"keywords":["Biology","Control of chromosome duplication","Replisome","DNA replication","Pre-replication complex","Eukaryotic DNA replication","Telomere","Origin recognition complex","Replication factor C","Telomere-binding protein","Minichromosome maintenance","S phase","Cell biology","Helicase","DNA","Genetics","DNA-binding protein","Gene","RNA","Transcription factor","570","Base Composition","Saccharomyces cerevisiae Proteins","Telomere-Binding Proteins","DNA Helicases","500","Gene Expression","Genome Integrity, Repair and Replication","In Vitro Techniques","Recombinant Proteins","Shelterin Complex","Saccharomycetales","Transcription Factors"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. Good health"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-11T02:24:44.109871Z","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":[]}