{"doi":"10.1016/j.jbc.2024.105753","title":"Monoubiquitination empowers ubiquitin chain elongation","abstract":"Ubiquitination often generates lysine 48-linked polyubiquitin chains that signal proteolytic destruction of the protein target. A significant subset of ubiquitination proceeds by a priming/extending mechanism, in which a substrate is first mono-ubiquitinated with a priming E2 conjugating enzyme or a set of E3 ARIH/E2 enzymes specific for priming. This is then followed by ubiquitin chain extension catalyzed by an E2 enzyme capable of elongation. This report provides further insights into the priming/extending mechanism. We employed reconstituted ubiquitination systems of substrates CK1α and β-catenin by Cullin-RING E3 ubiquitin ligases (CRLs) CRL4 CRBN and CRL1 βTrCP , respectively, in the presence of priming E2 UbcH5c and elongating E2 Cdc34b. We have established a new \"Apyrase chase\" strategy that uncouples priming from chain elongation, which allows accurate measurement of the decay rates of the ubiquitinated substrate with a defined chain length. Our work has revealed highly robust turnover of mono-ubiquitinated β-catenin that empowers efficient polyubiquitination. The results of competition experiments suggest that the interactions between the ubiquitinated β-catenin and CRL1 βTrCP are highly dynamic. Moreover, ubiquitination of the ubiquitin-modified β-catenin appeared more resistant to inhibition by competitors than the unmodified substrate, suggesting tighter binding with CRL1 βTrCP . These findings support a role for conjugated ubiquitin in enhancing interactions with E3.","journal":"Journal of Biological Chemistry","year":2024,"id":459200,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9478,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":469443,"name":"Robert J. DeVita","orcid":"0000-0002-2671-8497","position":1,"is_corresponding":false},{"id":383458,"name":"Zhen‐Qiang Pan","orcid":"0000-0001-9504-8768","position":2,"is_corresponding":false},{"id":383454,"name":"Kenneth Wu","orcid":"0000-0003-3513-5143","position":0,"is_corresponding":true}],"reference_count":31,"raw_metadata":null,"created_at":"2026-07-19T02:03:55.280882Z","pmid":"38354782","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":[]}