{"doi":"10.1101/2025.07.19.665708","title":"CDK1-dependent N-terminal NuMA phosphorylation promotes dynein-dynactin-NuMA assembly for accurate chromosome segregation","abstract":"Abstract The microtubule-based motor dynein and its cofactor dynactin fulfil essential functions throughout the cell cycle, including organelle transport and mitotic spindle assembly. To achieve these diverse functions, dynein-dynactin associates with different activating adaptors. Nuclear Mitotic Apparatus (NuMA) is a mitosis-specific adaptor that connects dynein-dynactin with microtubules to focus mitotic spindle poles. NuMA’s C-terminal microtubule-binding activity is promoted by mitotic phosphorylation, but regulation of NuMA’s N-terminal interaction with dynein-dynactin remains unclear. Here, we combine a membrane-tethering assay, quantitative proteomics, and live functional analyses in human cells to show that the interaction between NuMA’s N-terminus and dynein-dynactin is cell cycle-regulated and driven by mitotic phosphorylation. We identify highly conserved CDK1 consensus sites proximal to NuMA’s dynein heavy chain-binding site, which are phosphorylated by CDK1-Cyclin B1 in cells and in vitro . This CDK1-dependent phosphorylation, together with NuMA’s Spindly-like motif, is crucial for stable dynein-dynactin-NuMA (DDN) complex formation. Replacement of endogenous NuMA with phosphorylation-deficient NuMA mutants leads to aberrant dynein distribution on mitotic spindles, resulting in chromosome mis-segregation and micronucleus formation. Together, our results highlight CDK1-dependent N-terminal NuMA phosphorylation as a crucial mitotic switch that constitutes a regulatable core of multivalent interactions with dynein-dynactin to assemble stable DDN complexes for accurate chromosome segregation.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":558536,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9512,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1458948,"name":"Keishi Shintomi","orcid":"0000-0003-0484-9901","position":1,"is_corresponding":false},{"id":1458949,"name":"Tomomi Kiyomitsu","orcid":"0000-0002-2280-4611","position":2,"is_corresponding":false},{"id":1458947,"name":"Marvin van Toorn","orcid":"0000-0002-3992-9090","position":0,"is_corresponding":true}],"reference_count":83,"raw_metadata":null,"created_at":"2026-07-19T02:55:25.969263Z","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":[]}