{"doi":"10.1101/gad.250985.114","title":"Reconstitution of CPSF active in polyadenylation: recognition of the polyadenylation signal by WDR33","abstract":"<jats:p>Cleavage and polyadenylation specificity factor (CPSF) is the central component of the 3′ processing machinery for polyadenylated mRNAs in metazoans: CPSF recognizes the polyadenylation signal AAUAAA, providing sequence specificity in both pre-mRNA cleavage and polyadenylation, and catalyzes pre-mRNA cleavage. Here we show that of the seven polypeptides that have been proposed to constitute CPSF, only four (CPSF160, CPSF30, hFip1, and WDR33) are necessary and sufficient to reconstitute a CPSF subcomplex active in AAUAAA-dependent polyadenylation, whereas CPSF100, CPSF73, and symplekin are dispensable. WDR33 is required for binding of reconstituted CPSF to AAUAAA-containing RNA and can be specifically UV cross-linked to such RNAs, as can CPSF30. Transcriptome-wide identification of WDR33 targets by photoactivatable ribonucleoside-enhanced cross-linking and immunoprecipitation (PAR-CLIP) showed that WDR33 binds in and very close to the AAUAAA signal in vivo with high specificity. Thus, our data indicate that the large CPSF subunit participating in recognition of the polyadenylation signal is WDR33 and not CPSF160, as suggested by previous studies.</jats:p>","journal":"Genes &amp; Development","year":2014,"id":592910,"datarank":7.406930361057237,"base_score":5.424950017481403,"endowment":5.424950017481403,"self_citation_contribution":0.8137425026222106,"citation_network_contribution":6.593187858435026,"self_endowment_contribution":0.8137425026222106,"citer_contribution":6.593187858435026,"corpus_percentile":null,"corpus_rank":null,"citation_count":226,"citer_count":200,"citers_with_citation_signal":175,"citers_with_endowment":175,"datacite_reuse_total":18,"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":1517328,"name":"Uwe Kühn","orcid":null,"position":1,"is_corresponding":false},{"id":1509539,"name":"Georges Martin","orcid":null,"position":2,"is_corresponding":false},{"id":350497,"name":"Peter Schäfer","orcid":"0000-0002-3880-0850","position":3,"is_corresponding":false},{"id":1509538,"name":"Andreas R. Gruber","orcid":null,"position":4,"is_corresponding":false},{"id":79778,"name":"Walter Keller","orcid":"0000-0002-2261-958X","position":5,"is_corresponding":false},{"id":123393,"name":"Mihaela Zavolan","orcid":null,"position":6,"is_corresponding":false},{"id":308775,"name":"Elmar Wahle","orcid":"0000-0003-2504-0677","position":7,"is_corresponding":false},{"id":1517326,"name":"Lars Schönemann","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Reconstitution of CPSF active in polyadenylation: recognition of the polyadenylation signal by WDR33","abstract":"<jats:p>Cleavage and polyadenylation specificity factor (CPSF) is the central component of the 3′ processing machinery for polyadenylated mRNAs in metazoans: CPSF recognizes the polyadenylation signal AAUAAA, providing sequence specificity in both pre-mRNA cleavage and polyadenylation, and catalyzes pre-mRNA cleavage. Here we show that of the seven polypeptides that have been proposed to constitute CPSF, only four (CPSF160, CPSF30, hFip1, and WDR33) are necessary and sufficient to reconstitute a CPSF subcomplex active in AAUAAA-dependent polyadenylation, whereas CPSF100, CPSF73, and symplekin are dispensable. WDR33 is required for binding of reconstituted CPSF to AAUAAA-containing RNA and can be specifically UV cross-linked to such RNAs, as can CPSF30. Transcriptome-wide identification of WDR33 targets by photoactivatable ribonucleoside-enhanced cross-linking and immunoprecipitation (PAR-CLIP) showed that WDR33 binds in and very close to the AAUAAA signal in vivo with high specificity. Thus, our data indicate that the large CPSF subunit participating in recognition of the polyadenylation signal is WDR33 and not CPSF160, as suggested by previous studies.</jats:p>","is_dataset_classified":null,"base_score":5.424950017481403,"endowment":5.424950017481403,"datacite_reuse_total":18,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"25301781","pmcid":"PMC4215183","openalex_id":"https://openalex.org/W2166494447","authors":[],"funders":[{"funder_name":"Deutsche Forschungsgemeinschaft","grant_id":"WA 548/15-1","title":null},{"funder_name":"Swiss National Science Foundation","grant_id":"31003A-143977","title":null},{"funder_name":"Swiss National Science Foundation","grant_id":"143977","title":"The molecular biology of RNA 3' end processing"},{"funder_name":"Deutsche Forschungsgemeinschaft","grant_id":"unidentified","title":"unidentified"}],"total_grants":4,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[{"year":2014,"count":1},{"year":2015,"count":14},{"year":2016,"count":15},{"year":2017,"count":18},{"year":2018,"count":19},{"year":2019,"count":26},{"year":2020,"count":18},{"year":2021,"count":39},{"year":2022,"count":23},{"year":2023,"count":21},{"year":2024,"count":16},{"year":2025,"count":14},{"year":2026,"count":1}],"oa_status":"gold","license":"cc-by-nc","oa_locations":[{"url":"https://doi.org/10.1101/gad.250985.114","host_type":"journal"},{"url":"https://doi.org/10.1101/gad.250985.114","host_type":"publisher"},{"url":"https://syndication.highwire.org/content/doi/10.1101/gad.250985.114","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/25301781","host_type":"repository"},{"url":"http://genesdev.cshlp.org/cgi/content/short/28/21/2381","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/4215183","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC4215183","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC4215183?pdf=render","host_type":"Europe_PMC"},{"url":"https://dx.doi.org/10.5451/unibas-ep34873","host_type":""},{"url":"http://dx.doi.org/10.1101/gad.250985.114","host_type":""},{"url":"https://dx.doi.org/10.1101/gad.250985.114","host_type":""},{"url":"https://sonar.ch/global/documents/124314","host_type":""},{"url":"http://genesdev.cshlp.org/content/28/21/2381.full.html","host_type":""},{"url":"https://doi.org/https://doi.org/10.1101/gad.250985.114","host_type":""}],"fields_of_study":["RNA Research and Splicing","RNA modifications and cancer","Cancer-related gene regulation","0301 basic medicine","0303 health sciences","03 medical and health sciences","Cleavage And Polyadenylation Specificity Factor","Gene Expression Profiling","Gene Expression Regulation","HEK293 Cells","Humans","Nuclear Proteins","Polyadenylation","Protein Binding","Protein Subunits","RNA 3' End Processing"],"mesh_terms":["Gene Expression Regulation","Humans","Nuclear Proteins","Protein Binding","Gene Expression Profiling","Protein Subunits","RNA 3' End Processing","Polyadenylation","Cleavage And Polyadenylation Specificity Factor","HEK293 Cells"],"keywords":["Cleavage and polyadenylation specificity factor","Polyadenylation","Biology","Cleavage factor","Cleavage stimulation factor","Post-transcriptional modification","RNA-binding protein","Cell biology","Cleavage (geology)","Messenger RNA","Biochemistry","Gene","RNA processing","Poly(a) Polymerase","3′ End Formation","Poly(a) Site","Gene Expression Profiling","Nuclear Proteins","Protein Subunits","HEK293 Cells","Gene Expression Regulation","Humans","RNA 3' End Processing","Research Paper","Protein Binding"],"sdg_mappings":[],"linked_datasets":[{"doi":"10.6084/m9.figshare.14205366.v1","title":"Additional file 10 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205366","title":"Additional file 10 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205372.v1","title":"Additional file 11 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205372","title":"Additional file 11 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205375.v1","title":"Additional file 1 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205375","title":"Additional file 1 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205402.v1","title":"Additional file 9 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14205402","title":"Additional file 9 of SRSF3 and SRSF7 modulate 3′UTR length through suppression or activation of proximal polyadenylation sites and regulation of CFIm levels","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853530.v1","title":"Additional file 1 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853530","title":"Additional file 1 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853533.v1","title":"Additional file 2 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853533","title":"Additional file 2 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853536.v1","title":"Additional file 3 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853536","title":"Additional file 3 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853539.v1","title":"Additional file 4 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.14853539","title":"Additional file 4 of LABRAT reveals association of alternative polyadenylation with transcript localization, RNA binding protein expression, transcription speed, and cancer survival","publisher":"figshare","resource_type":"JournalArticle"},{"doi":"10.6084/m9.figshare.13545670","title":"Additional file 1 of Long non-coding RNA FAM83H-AS1 acts as a potential oncogenic driver in human ovarian cancer","publisher":"figshare","resource_type":"Dataset"},{"doi":"10.6084/m9.figshare.13545670.v1","title":"Additional file 1 of Long non-coding RNA FAM83H-AS1 acts as a potential oncogenic driver in human ovarian cancer","publisher":"figshare","resource_type":"Dataset"}],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"geo"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-26T16:09:55.900658Z","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":[]}