title: GSK-3β Phosphorylation of Cytoplasmic Dynein Reduces Ndel1 Binding to Intermediate Chains and Alters Dynein Motility.
abstract: Glycogen synthase kinase 3 (GSK-3) has been linked to regulation of kinesin-dependent axonal transport in squid and flies, and to indirect regulation of cytoplasmic dynein. We have now found evidence for direct regulation of dynein by mammalian GSK-3β in both neurons and non-neuronal cells. GSK-3β coprecipitates with and phosphorylates mammalian dynein. Phosphorylation of dynein intermediate chain (IC) reduces its interaction with Ndel1, a protein that contributes to dynein force generation. Two conserved residues, S87/T88 in IC-1B and S88/T89 in IC-2C, have been identified as GSK-3 targets by both mass spectrometry and site-directed mutagenesis. These sites are within an Ndel1-binding domain, and mutation of both sites alters the interaction of IC's with Ndel1. Dynein motility is stimulated by (i) pharmacological and genetic inhibition of GSK-3β, (ii) an insulin-sensitizing agent (rosiglitazone) and (iii) manipulating an insulin response pathway that leads to GSK-3β inactivation. Thus, our study connects a well-characterized insulin-signaling pathway directly to dynein stimulation via GSK-3 inhibition.
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: Feng J
authors.initials: FJ
authors.lastname: Gao
authors.affiliation: Bioinformatics Group, Immune Tolerance Network, Bethesda, MD, 20814, USA.
authors.forename: Sachin
authors.initials: S
authors.lastname: Hebbar
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: Xu A
authors.initials: XA
authors.lastname: Gao
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Alexander
authors.affiliation: Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA, 02142, USA.
authors.forename: Jai P
authors.initials: JP
authors.lastname: Pandey
authors.affiliation: Mass Spectrometry Center, Department of Chemistry & Biochemistry, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Walla
authors.affiliation: Mass Spectrometry Center, Department of Chemistry & Biochemistry, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: William E
authors.initials: WE
authors.lastname: Cotham
authors.affiliation: Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, 32828, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: King
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC, 29208, USA.
authors.forename: Deanna S
authors.initials: DS
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D004398:Dyneins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D021381:Protein Transport; D015290:Second Messenger Systems
chemical_list: D002352:Carrier Proteins; D007328:Insulin; C486015:Ndel1 protein, mouse; D038362:Glycogen Synthase Kinase 3; D004398:Dyneins
keywords: Ndel1/Nde1; axon transport; centrosome; cytoplasmic dynein; insulin; retrograde transport