{"doi":"10.1038/ncomms14286","title":"Orthogonal ubiquitin transfer identifies ubiquitination substrates under differential control by the two ubiquitin activating enzymes","abstract":"<jats:title>Abstract</jats:title><jats:p>Protein ubiquitination is mediated sequentially by ubiquitin activating enzyme E1, ubiquitin conjugating enzyme E2 and ubiquitin ligase E3. Uba1 was thought to be the only E1 until the recent identification of Uba6. To differentiate the biological functions of Uba1 and Uba6, we applied an orthogonal ubiquitin transfer (OUT) technology to profile their ubiquitination targets in mammalian cells. By expressing pairs of an engineered ubiquitin and engineered Uba1 or Uba6 that were generated for exclusive interactions, we identified 697 potential Uba6 targets and 527 potential Uba1 targets with 258 overlaps. Bioinformatics analysis reveals substantial differences in pathways involving Uba1- and Uba6-specific targets. We demonstrate that polyubiquitination and proteasomal degradation of ezrin and CUGBP1 require Uba6, but not Uba1, and that Uba6 is involved in the control of ezrin localization and epithelial morphogenesis. These data suggest that distinctive substrate pools exist for Uba1 and Uba6 that reflect non-redundant biological roles for Uba6.</jats:p>","journal":"Nature Communications","year":2017,"id":24310,"datarank":1.5950486790943548,"base_score":3.8712010109078907,"endowment":3.8712010109078907,"self_citation_contribution":0.5806801516361837,"citation_network_contribution":1.014368527458171,"self_endowment_contribution":0.5806801516361837,"citer_contribution":1.014368527458171,"corpus_percentile":null,"corpus_rank":null,"citation_count":47,"citer_count":36,"citers_with_citation_signal":32,"citers_with_endowment":32,"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":36210,"name":"Bo Zhao","orcid":"0000-0001-7188-5959","position":1,"is_corresponding":false},{"id":146284,"name":"Limin Sun","orcid":null,"position":2,"is_corresponding":false},{"id":36212,"name":"Karan Bhuripanyo","orcid":"0000-0002-0945-6085","position":3,"is_corresponding":false},{"id":146285,"name":"Yiyang Wang","orcid":null,"position":4,"is_corresponding":false},{"id":90579,"name":"Yingtao Bi","orcid":"0000-0003-4088-6558","position":5,"is_corresponding":false},{"id":89831,"name":"Ramana V. Davuluri","orcid":"0000-0002-7053-1064","position":6,"is_corresponding":false},{"id":21522,"name":"Duc M. Duong","orcid":"0000-0001-5880-1888","position":7,"is_corresponding":false},{"id":146286,"name":"Dhaval Nanavati","orcid":null,"position":8,"is_corresponding":false},{"id":36217,"name":"Jun Yin","orcid":"0000-0002-4803-7510","position":9,"is_corresponding":false},{"id":36215,"name":"Hiroaki Kiyokawa","orcid":"0000-0002-7942-6455","position":10,"is_corresponding":false},{"id":146283,"name":"Xianpeng Liu","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.8712010109078907,"endowment":3.8712010109078907,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"28134249","pmcid":"PMC5290280","openalex_id":"https://openalex.org/W2579904589","authors":[],"funders":[{"funder_name":"NCI NIH HHS","grant_id":"P30 CA010815","title":null},{"funder_name":"NLM NIH HHS","grant_id":"R01 LM011297","title":null},{"funder_name":"NCI NIH HHS","grant_id":"P30 CA060553","title":null},{"funder_name":"NCI NIH HHS","grant_id":"R01 CA112282","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM104498","title":null},{"funder_name":"National Science Foundation","grant_id":"1057092","title":"CAREER: Identifying the Substrate Proteins of Nedd4 Family of E3 Ubiquitin Ligase by Orthogonal Ubiquitin Transfer (OUT)"},{"funder_name":"National Institutes of Health","grant_id":"3R01GM104498-08S1","title":"Orthogonal Ubiquitin Transfer to Profile E3 Substrate Specificity."},{"funder_name":"National Institutes of Health","grant_id":"2R01CA112282-06A1","title":"Role of Cdc25A in breast cancer"}],"total_grants":8,"fwci":2.4668,"citation_percentile":0.8948406,"influential_citations":1,"citation_trend":[{"year":2017,"count":2},{"year":2018,"count":7},{"year":2019,"count":5},{"year":2020,"count":8},{"year":2021,"count":5},{"year":2022,"count":6},{"year":2023,"count":2},{"year":2024,"count":8},{"year":2025,"count":3},{"year":2026,"count":1}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.nature.com/articles/ncomms14286.pdf","host_type":"journal"},{"url":"https://doi.org/10.1038/ncomms14286","host_type":"GOLD"},{"url":"https://www.nature.com/articles/ncomms14286.pdf","host_type":"publisher"},{"url":"https://www.nature.com/articles/ncomms14286","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/28134249","host_type":"repository"},{"url":"https://doaj.org/article/770ad999b01440b0979cab03270cc849","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/5290280","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC5290280","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC5290280?pdf=render","host_type":"Europe_PMC"},{"url":"http://dx.doi.org/10.1038/ncomms14286","host_type":""},{"url":"https://dx.doi.org/10.1038/ncomms14286","host_type":""}],"fields_of_study":["Ubiquitin and proteasome pathways","Glycosylation and Glycoproteins Research","Cellular transport and secretion","Biology","Medicine","Chemistry","0301 basic medicine","03 medical and health sciences","0303 health sciences","CELF1 Protein","Cell Differentiation","Cell Line, Tumor","Computational Biology","Cytoskeletal Proteins","Epithelium","HEK293 Cells","Humans","Morphogenesis","Mutagenesis, Insertional","Proteasome Endopeptidase Complex","Proteolysis","RNA, Small Interfering","Signal Transduction","Substrate Specificity","Ubiquitin","Ubiquitin-Activating Enzymes","Ubiquitination","Ezrin"],"mesh_terms":["CELF1 Protein","Ezrin","Cell Differentiation","Cytoskeletal Proteins","Epithelium","Humans","Morphogenesis","Substrate Specificity","Signal Transduction","Mutagenesis, Insertional","Computational Biology","Ubiquitin","RNA, Small Interfering","Ubiquitin-Activating Enzymes","Cell Line, Tumor","Proteasome Endopeptidase Complex","Ubiquitination","HEK293 Cells","Proteolysis"],"keywords":["Ubiquitin","Ubiquitin ligase","Ubiquitin-conjugating enzyme","Ubiquitin-Protein Ligases","Deubiquitinating enzyme","Cell biology","Ubiquitins","Biology","Ezrin","Computational biology","Biochemistry","Chemistry","Gene","Cell","Proteasome Endopeptidase Complex","Science","Ubiquitin-Activating Enzymes","Article","Epithelium","Substrate Specificity","Cell Line, Tumor","Morphogenesis","Humans","RNA, Small Interfering","CELF1 Protein","Q","Ubiquitination","Cell Differentiation","Cytoskeletal Proteins","Mutagenesis, Insertional","HEK293 Cells","Proteolysis","Signal Transduction"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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