{"doi":"10.1186/1471-2091-12-18","title":"Effect of reverse transcriptase inhibitors on LINE-1 and Ty1 reverse transcriptase activities and on LINE-1 retrotransposition","abstract":"<jats:title>Abstract</jats:title>\n          <jats:sec>\n            <jats:title>Background</jats:title>\n            <jats:p>LINE-1s (L1, Long Interspersed Element-1) are the most abundant autonomous non-LTR retrotransposons in the human genome and replicate by reverse transcription of an RNA intermediate. Full-length L1 encodes two open reading frames (ORF1, ORF2) and ORF2 has reverse transcriptase activity.</jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>Results</jats:title>\n            <jats:p>Here we expressed human L1 RT in <jats:italic>E. coli</jats:italic> and the purified protein displayed the same RT activity as that of ORF2p expressed in insect cells. We tested the effect of different reverse transcriptase inhibitors on L1 RT and found that all four tested nucleoside inhibitors efficiently inhibited L1 RT activity competitively. The K<jats:sub>i</jats:sub> values of NRTIs were calculated (AZTTP, 16.4 ± 4.21 nM; d4TTP, 0.73 ± 0.22 nM; ddCTP, 0.72 ± 0.16 nM; 3TCTP, 12.9 ± 2.07 nM). L1 RT was less sensitive to non-nucleoside reverse transcriptase inhibitors, among these nevirapine had no effect, even at concentrations up to 500 μM. We also examined the effect of RT inhibitors on L1 retrotransposition efficiency <jats:italic>in vivo</jats:italic> using a cell-based retrotransposition assay. Similarly, all analog inhibitors decreased L1 retrotransposition frequency with different potencies whereas nevirapine had little or no effect on L1 retrotransposition. For comparison, we also tested the same inhibitors to highly purified RT of an LTR-retrotransposon (Ty1) and found it was less sensitive to NRTIs than L1 RT and has the same inhibition profile as L1 RT to NNRTIs.</jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions</jats:title>\n            <jats:p>These data indicate that bacterially expressed L1 RT is an active reverse transcriptase sensitive to nucleoside RT inhibitors but not to non-nucleoside inhibitors.</jats:p>\n          </jats:sec>","journal":"BMC Biochemistry","year":2011,"id":612355,"datarank":0.7379971388742189,"base_score":4.919980925828125,"endowment":4.919980925828125,"self_citation_contribution":0.7379971388742189,"citation_network_contribution":0.0,"self_endowment_contribution":0.7379971388742189,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":136,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":6,"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":122781,"name":"Qing Huang","orcid":null,"position":1,"is_corresponding":false},{"id":114985,"name":"Jef D Boeke","orcid":null,"position":2,"is_corresponding":false},{"id":1393959,"name":"Lixin Dai","orcid":"0000-0001-5090-5180","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Effect of reverse transcriptase inhibitors on LINE-1 and Ty1 reverse transcriptase activities and on LINE-1 retrotransposition","abstract":"<jats:title>Abstract</jats:title>\n          <jats:sec>\n            <jats:title>Background</jats:title>\n            <jats:p>LINE-1s (L1, Long Interspersed Element-1) are the most abundant autonomous non-LTR retrotransposons in the human genome and replicate by reverse transcription of an RNA intermediate. Full-length L1 encodes two open reading frames (ORF1, ORF2) and ORF2 has reverse transcriptase activity.</jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>Results</jats:title>\n            <jats:p>Here we expressed human L1 RT in <jats:italic>E. coli</jats:italic> and the purified protein displayed the same RT activity as that of ORF2p expressed in insect cells. We tested the effect of different reverse transcriptase inhibitors on L1 RT and found that all four tested nucleoside inhibitors efficiently inhibited L1 RT activity competitively. The K<jats:sub>i</jats:sub> values of NRTIs were calculated (AZTTP, 16.4 ± 4.21 nM; d4TTP, 0.73 ± 0.22 nM; ddCTP, 0.72 ± 0.16 nM; 3TCTP, 12.9 ± 2.07 nM). L1 RT was less sensitive to non-nucleoside reverse transcriptase inhibitors, among these nevirapine had no effect, even at concentrations up to 500 μM. We also examined the effect of RT inhibitors on L1 retrotransposition efficiency <jats:italic>in vivo</jats:italic> using a cell-based retrotransposition assay. Similarly, all analog inhibitors decreased L1 retrotransposition frequency with different potencies whereas nevirapine had little or no effect on L1 retrotransposition. For comparison, we also tested the same inhibitors to highly purified RT of an LTR-retrotransposon (Ty1) and found it was less sensitive to NRTIs than L1 RT and has the same inhibition profile as L1 RT to NNRTIs.</jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions</jats:title>\n            <jats:p>These data indicate that bacterially expressed L1 RT is an active reverse transcriptase sensitive to nucleoside RT inhibitors but not to non-nucleoside inhibitors.</jats:p>\n          </jats:sec>","is_dataset_classified":null,"base_score":4.919980925828125,"endowment":4.919980925828125,"datacite_reuse_total":6,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"21545744","pmcid":"PMC3103432","openalex_id":"https://openalex.org/W1982644994","authors":[],"funders":[{"funder_name":"NCI NIH HHS","grant_id":"P01-CA16519","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"R01-GM36481","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM036481","title":null},{"funder_name":"National Institutes of Health","grant_id":"5R01CA036481-19","title":"ABERRANT REGULATION OF CATHEPSIN B EXPRESSION IN TUMORS"},{"funder_name":"Fundação para a Ciência e a Tecnologia, I.P.","grant_id":"PTDC/CCI-BIO/29266/2017","title":"Deep Drug Discovery and Deployment"},{"funder_name":"National Institutes of Health","grant_id":"5P01CA016519-19","title":"PROGRAM ON MOLECULAR BIOLOGY OF VIRAL TUMORIGENESIS"}],"total_grants":6,"fwci":7.1002,"citation_percentile":0.96626605,"influential_citations":0,"citation_trend":[{"year":2012,"count":1},{"year":2013,"count":11},{"year":2014,"count":6},{"year":2015,"count":6},{"year":2016,"count":4},{"year":2017,"count":11},{"year":2018,"count":6},{"year":2019,"count":13},{"year":2020,"count":18},{"year":2021,"count":11},{"year":2022,"count":17},{"year":2023,"count":7},{"year":2024,"count":8},{"year":2025,"count":11},{"year":2026,"count":5}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"https://bmcbiochem.biomedcentral.com/counter/pdf/10.1186/1471-2091-12-18","host_type":"journal"},{"url":"https://bmcbiochem.biomedcentral.com/counter/pdf/10.1186/1471-2091-12-18","host_type":"publisher"},{"url":"https://link.springer.com/content/pdf/10.1186/1471-2091-12-18.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1186/1471-2091-12-18","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/21545744","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/3103432","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC3103432","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC3103432?pdf=render","host_type":"Europe_PMC"},{"url":"https://bmcbiochem.biomedcentral.com/track/pdf/10.1186/1471-2091-12-18","host_type":""},{"url":"http://dx.doi.org/10.1186/1471-2091-12-18","host_type":""},{"url":"https://dx.doi.org/10.1186/1471-2091-12-18","host_type":""}],"fields_of_study":["Chromosomal and Genetic Variations","Genomic variations and chromosomal abnormalities","Nuclear Structure and Function","0301 basic medicine","0303 health sciences","03 medical and health sciences","Amino Acid Sequence","Cell-Free System","Enzyme Assays","Escherichia coli","HIV-1","HIV-2","HeLa Cells","Humans","Kinetics","Long Interspersed Nucleotide Elements","Molecular Sequence Data","RNA-Directed DNA Polymerase","Reverse Transcriptase Inhibitors"],"mesh_terms":["Amino Acid Sequence","Cell-Free System","Escherichia coli","HeLa Cells","Humans","Kinetics","Molecular Sequence Data","RNA-Directed DNA Polymerase","HIV-1","HIV-2","Reverse Transcriptase Inhibitors","Long Interspersed Nucleotide Elements","Enzyme Assays","Hela Cells"],"keywords":["Retrotransposon","Reverse transcriptase","Nevirapine","Nucleoside Reverse Transcriptase Inhibitor","Biology","RNA","Nucleotidyltransferase","Molecular biology","Reverse-transcriptase inhibitor","Long terminal repeat","Nucleoside","Virology","Biochemistry","Human immunodeficiency virus (HIV)","Genome","Gene","Transposable element","Cell-Free System","Molecular Sequence Data","RNA-Directed DNA Polymerase","Kinetics","Long Interspersed Nucleotide Elements","HIV-2","Escherichia coli","HIV-1","Humans","Reverse Transcriptase Inhibitors","Amino Acid Sequence","Research Article","Enzyme Assays","HeLa Cells"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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