{"doi":"10.1101/2021.10.12.463902","title":"The mechanochemical origins of the microtubule sliding motility within the kinesin-5 domain organization","abstract":"Abstract The conserved kinesin-5 bipolar tetrameric motors slide apart microtubules during mitotic spindle assembly and elongation. Kinesin-5 bipolar organization originates from its conserved tetrameric helical minifilament, which position the C-terminal tail domains of two subunits near the N-terminal motor domains of two anti-parallel subunits (Scholey et al, 2014). This unique tetrameric structure enables kinesin-5 to simultaneously engage two microtubules and transmit forces between them, and for multiple kinesin-5 motors to organize via tail to motor interactions during microtubule sliding (Bodrug et al, 2020). Here, we show how these two structural adaptations, the kinesin-5 tail-motor domain interactions and the length of the tetrameric minifilament, determine critical aspects of kinesin-5 motility and sliding mechanisms. An x-ray structure of the 34-nm kinesin-5 minifilament reveals how the dual dimeric N-terminal coiled-coils emerge from the tetrameric central bundle. Using this structure, we generated active bipolar mini-tetrameric motors from Drosophila and human orthologs, which are half the length of native kinesin-5. Using single-molecule motility assays, we show that kinesin-5 tail domains promote mini-tetramers static pauses that punctuate processive motility. During such pauses, kinesin-5 mini-tetramers form multi-motor clusters mediated via tail to motor domain cross-interactions. These clusters undergo slow and highly processive motility and accumulate at microtubule plus-ends. In contrast to native kinesin-5, mini-tetramers require tail domains to initiate microtubule crosslinking. Although mini-tetramers are highly strained in initially aligning microtubules, they slide microtubules more efficiently than native kinesin-5, due to their decreased minifilament flexibility. Our studies reveal that the conserved kinesin-5 motor-tail mediated clustering and the length of the tetrameric minifilament are key features for sliding motility and are critical in organizing microtubules during mitotic spindle assembly and elongation.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2021,"id":217951,"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":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9498,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":815286,"name":"Malina K. Iwanski","orcid":"0000-0002-4903-9796","position":1,"is_corresponding":false},{"id":326786,"name":"Ignas Gaska","orcid":"0009-0005-2569-0751","position":2,"is_corresponding":false},{"id":566263,"name":"Himanshu Pandey","orcid":"0000-0002-0629-7525","position":3,"is_corresponding":false},{"id":241647,"name":"Tatyana Bodrug","orcid":"0000-0001-9017-962X","position":4,"is_corresponding":false},{"id":326788,"name":"Sayaka Inagaki","orcid":"0000-0001-9064-0762","position":5,"is_corresponding":false},{"id":326787,"name":"Jennifer Major","orcid":"0000-0003-3140-5046","position":6,"is_corresponding":false},{"id":556388,"name":"Gary J. Brouhard","orcid":"0000-0001-9101-1247","position":7,"is_corresponding":false},{"id":326790,"name":"Larisa Gheber","orcid":"0000-0003-3759-4001","position":8,"is_corresponding":false},{"id":326792,"name":"Steven S. Rosenfeld","orcid":"0000-0003-1909-7116","position":9,"is_corresponding":false},{"id":326793,"name":"Scott Forth","orcid":"0000-0002-2630-8017","position":10,"is_corresponding":false},{"id":815249,"name":"Adam G. Hendricks","orcid":"0000-0003-3635-1577","position":11,"is_corresponding":false},{"id":326794,"name":"Jawdat Al‐Bassam","orcid":"0000-0001-6625-2102","position":12,"is_corresponding":false},{"id":326784,"name":"Stanley Nithianantham","orcid":"0000-0001-6238-647X","position":0,"is_corresponding":true}],"reference_count":52,"raw_metadata":null,"created_at":"2026-07-18T23:53:24.683898Z","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":[]}