title: The N-terminal coiled-coil of Ndel1 is a regulated scaffold that recruits LIS1 to dynein.
abstract: Ndel1 has been implicated in a variety of dynein-related processes, but its specific function is unclear. Here we describe an experimental approach to evaluate a role of Ndel1 in dynein-dependent microtubule self-organization using Ran-mediated asters in meiotic Xenopus egg extracts. We demonstrate that extracts depleted of Ndel1 are unable to form asters and that this defect can be rescued by the addition of recombinant N-terminal coiled-coil domain of Ndel1. Ndel1-dependent microtubule self-organization requires an interaction between Ndel1 and dynein, which is mediated by the dimerization fragment of the coiled-coil. Full rescue by the coiled-coil domain requires LIS1 binding, and increasing LIS1 concentration partly rescues aster formation, suggesting that Ndel1 is a recruitment factor for LIS1. The interactions between Ndel1 and its binding partners are positively regulated by phosphorylation of the unstructured C terminus. Together, our results provide important insights into how Ndel1 acts as a regulated scaffold to temporally and spatially regulate dynein.
authors.affiliation: Department of Biochemistry, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
authors.forename: Eliza
authors.initials: E
authors.lastname: Zyłkiewicz
authors.forename: Monika
authors.initials: M
authors.lastname: Kijańska
authors.forename: Won-Chan
authors.initials: WC
authors.lastname: Choi
authors.forename: Urszula
authors.initials: U
authors.lastname: Derewenda
authors.forename: Zygmunt S
authors.initials: ZS
authors.lastname: Derewenda
authors.forename: P Todd
authors.initials: PT
authors.lastname: Stukenberg
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004398:Dyneins / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014981:Xenopus; D029867:Xenopus Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; C558828:LIS1 protein, Xenopus; D008869:Microtubule-Associated Proteins; C555415:Ndel1 protein, Xenopus; C486015:Ndel1 protein, mouse; D009687:Nuclear Proteins; D029867:Xenopus Proteins; D004398:Dyneins