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How JIP3\u2019s interaction with dynein and kinesin-1 contributes to organelle clearance is unclear. We show that human dynein light intermediate chain (DLIC) binds the N-terminal RH1 domain of JIP3, its paralog JIP4, and the lysosomal adaptor RILP. A point mutation in RH1 abrogates DLIC binding without perturbing the interaction between JIP3\u2019s RH1 domain and kinesin heavy chain. Characterization of this separation-of-function mutation in Caenorhabditis elegans shows that JIP3\u2013bound dynein is required for organelle clearance in the anterior process of touch receptor neurons. Unlike JIP3 null mutants, JIP3 that cannot bind DLIC causes prominent accumulation of endo-lysosomal organelles at the neurite tip, which is rescued by a disease-associated point mutation in JIP3\u2019s leucine zipper that abrogates kinesin light chain binding. These results highlight that RH1 domains are interaction hubs for cytoskeletal motors and suggest that JIP3\u2013bound dynein and kinesin-1 participate in bidirectional organelle transport.<\/jats:p>","DOI":"10.1083\/jcb.202110057","type":"journal-article","created":{"date-parts":[[2022,6,24]],"date-time":"2022-06-24T02:21:26Z","timestamp":1656037286000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":48,"title":["JIP3 interacts with dynein and kinesin-1 to regulate bidirectional organelle transport"],"prefix":"10.1083","volume":"221","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9605-6433","authenticated-orcid":true,"given":"Ricardo","family":"Celestino","sequence":"first","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7011-1502","authenticated-orcid":true,"given":"Jos\u00e9 B.","family":"Gama","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"given":"Artur F.","family":"Castro-Rodrigues","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1726-6011","authenticated-orcid":true,"given":"Daniel J.","family":"Barbosa","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"},{"name":"TOXRUN\u2014Toxicology Research Unit, University Institute of Health Sciences, Advanced Polytechnic and University Cooperative (CESPU), Cooperative of Limited Liability (CRL), Gandra, Portugal 2"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1062-2498","authenticated-orcid":true,"given":"Helder","family":"Rocha","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1451-6273","authenticated-orcid":true,"given":"Ennio A.","family":"d\u2019Amico","sequence":"additional","affiliation":[{"name":"Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany 3"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2362-8784","authenticated-orcid":true,"given":"Andrea","family":"Musacchio","sequence":"additional","affiliation":[{"name":"Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany 3"},{"name":"Centre for Medical Biotechnology, Faculty of Biology, University Duisburg-Essen, Essen, Germany 4"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2256-7134","authenticated-orcid":true,"given":"Ana Xavier","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4461-9716","authenticated-orcid":true,"given":"Jo\u00e3o H.","family":"Morais-Cabral","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0360-2977","authenticated-orcid":true,"given":"Reto","family":"Gassmann","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade\u2014i3S, Universidade do Porto, Porto, Portugal 1"}]}],"member":"291","published-online":{"date-parts":[[2022,7,13]]},"reference":[{"key":"2023072408510233600_bib1","doi-asserted-by":"publisher","first-page":"34628","DOI":"10.1074\/jbc.M109.035022","article-title":"Sunday driver interacts with two distinct classes of axonal organelles","volume":"284","author":"Abe","year":"2009","journal-title":"J. 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