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Here, we identify TBC1D20 as a novel Rab11 GTPase-activating protein that coordinates vesicle transport and actin remodeling to regulate ciliogenesis. Depletion of TBC1D20 promotes Rab11 vesicle accumulation and actin deconstruction around the centrosome, facilitating the initiation of ciliogenesis even in cycling cells. Further investigations reveal enhanced Rab11\u2013MICAL1 interaction upon TBC1D20 loss, activating the monooxygenase domain of MICAL1 and inducing F-actin depolymerization around the centrosome. This actin network reorganization facilitates vesicle trafficking and docking, ultimately promoting ciliogenesis. In summary, our work uncovers a coordinated ciliogenesis initiation mechanism via Rab11 activation. These findings underscore a pivotal role for TBC1D20 in early ciliogenesis, advancing our understanding of its spatiotemporal regulation and offering insights into the disease pathogenesis associated with TBC1D20 mutations.<\/jats:p>","DOI":"10.1083\/jcb.202406139","type":"journal-article","created":{"date-parts":[[2025,1,4]],"date-time":"2025-01-04T02:20:59Z","timestamp":1735957259000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":7,"title":["TBC1D20 coordinates vesicle transport and actin remodeling to regulate ciliogenesis"],"prefix":"10.1083","volume":"224","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-0994-1847","authenticated-orcid":true,"given":"Denghui","family":"Zhai","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01y1kjr75","id-type":"ROR","asserted-by":"publisher"}],"name":"Nankai University 1 Department of Genetics and Cell Biology, College of Life Sciences, State Key Laboratory of 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