title: Specificity of cytoplasmic dynein subunits in discrete membrane-trafficking steps.
abstract: The cytoplasmic dynein motor complex is known to exist in multiple forms, but few specific functions have been assigned to individual subunits. A key limitation in the analysis of dynein in intact mammalian cells has been the reliance on gross perturbation of dynein function, e.g., inhibitory antibodies, depolymerization of the entire microtubule network, or the use of expression of dominant negative proteins that inhibit dynein indirectly. Here, we have used RNAi and automated image analysis to define roles for dynein subunits in distinct membrane-trafficking processes. Depletion of a specific subset of dynein subunits, notably LIC1 (DYNC1LI1) but not LIC2 (DYNC1LI2), recapitulates a direct block of ER export, revealing that dynein is required to maintain the steady-state composition of the Golgi, through ongoing ER-to-Golgi transport. Suppression of LIC2 but not of LIC1 results in a defect in recycling endosome distribution and cytokinesis. Biochemical analyses also define the role of each subunit in stabilization of the dynein complex; notably, suppression of DHC1 or IC2 results in concomitant loss of Tctex1. Our data demonstrate that LIC1 and LIC2 define distinct dynein complexes that function at the Golgi versus recycling endosomes, respectively, suggesting that functional populations of dynein mediate discrete intracellular trafficking pathways.
authors.affiliation: Cell Biology Laboratories, Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS81TD, United Kingdom.
authors.forename: Krysten J
authors.initials: KJ
authors.lastname: Palmer
authors.forename: Helen
authors.initials: H
authors.lastname: Hughes
authors.forename: David J
authors.initials: DJ
authors.lastname: Stephens
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D003593:Cytoplasm / Q000378:metabolism*; D056445:Cytoplasmic Dyneins; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D020869:Gene Expression Profiling; D020868:Gene Silencing; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008870:Microtubules / Q000378:metabolism; D008954:Models, Biological; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D034741:RNA, Small Interfering; D014168:Transferrin; C424131:DYNC1LI1 protein, human; D056445:Cytoplasmic Dyneins; D004398:Dyneins