biostudies-literatureGirao H"la Caixa" Foundation (Caixa Foundation)Ministry of Education and Science | Fundação para a Ciência e a Tecnologia (Portuguese Science and Technology Foundation)European Research CouncilU.S. Department of Health & Human Services | National Institutes of Health (NIH)NIGMS NIH HHS9720https://www.ebi.ac.uk/biostudies/studies/S-EPMC11550433biostudies-literatureUnknown15(1)Post-translational cycles of α-tubulin detyrosination and tyrosination generate microtubule diversity, the cellular functions of which remain largely unknown. Here we show that α-tubulin detyrosination regulates kinetochore-microtubule attachments to ensure normal chromosome oscillations and timely anaphase onset during mitosis. Remarkably, detyrosinated α-tubulin levels near kinetochore microtubule plus-ends depend on the direction of chromosome motion during metaphase. Proteomic analyses unveil that the KNL-1/MIS12/NDC80 (KMN) network that forms the core microtubule-binding site at kinetochores and the microtubule-rescue protein CLASP2 are enriched on tyrosinated and detyrosinated microtubules during mitosis, respectively. α-tubulin detyrosination enhances CLASP2 binding and NDC80 compleNature communicationsα-tubulin detyrosination fine-tunes kinetochore-microtubule attachments.PMC11550433SFRH/BD/52044/2012SFRH/BD/141066/2018PTDC/MED-ONC/3479/2020R35 GM141747681443LCF/PR/HR21/524100252021.07945.BDR35-GM141747Meraldi PSilva E Sousa RMacario-Monteiro JDemidov VFigueiredo ACGirao HDoria EOsorio HJacome AMaiato HGrishchuk ELfalseα-tubulin detyrosination fine-tunes kinetochore-microtubule attachments.Post-translational cycles of α-tubulin detyrosination and tyrosination generate microtubule diversity, the cellular functions of which remain largely unknown. Here we show that α-tubulin detyrosination regulates kinetochore-microtubule attachments to ensure normal chromosome oscillations and timely anaphase onset during mitosis. Remarkably, detyrosinated α-tubulin levels near kinetochore microtubule plus-ends depend on the direction of chromosome motion during metaphase. Proteomic analyses unveil that the KNL-1/MIS12/NDC80 (KMN) network that forms the core microtubule-binding site at kinetochores and the microtubule-rescue protein CLASP2 are enriched on tyrosinated and detyrosinated microtubules during mitosis, respectively. α-tubulin detyrosination enhances CLASP2 binding and NDC80 comple2024-01-01T00:00:00Z2024 Nov2026-06-01T17:41:49.828Z2025-04-06T18:28:03.41ZS-EPMC115504333952180510.1038/s41467-024-54155-8