title: Reconstitution of dynein transport to the microtubule plus end by kinesin.
abstract: Cytoplasmic dynein powers intracellular movement of cargo toward the microtubule minus end. The first step in a variety of dynein transport events is the targeting of dynein to the dynamic microtubule plus end, but the molecular mechanism underlying this spatial regulation is not understood. Here, we reconstitute dynein plus-end transport using purified proteins from S. cerevisiae and dissect the mechanism using single-molecule microscopy. We find that two proteins-homologs of Lis1 and Clip170-are sufficient to couple dynein to Kip2, a plus-end-directed kinesin. Dynein is transported to the plus end by Kip2, but is not a passive passenger, resisting its own plus-end-directed motion. Two microtubule-associated proteins, homologs of Clip170 and EB1, act as processivity factors for Kip2, helping it overcome dynein's intrinsic minus-end-directed motility. This reveals how a minimal system of proteins transports a molecular motor to the start of its track.DOI: http://dx.doi.org/10.7554/eLife.02641.001.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, United States Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom anthony_roberts@hms.harvard.edu.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Roberts
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, United States.
authors.forename: Brian S
authors.initials: BS
authors.lastname: Goodman
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, United States reck-peterson@hms.harvard.edu.
authors.forename: Samara L
authors.initials: SL
authors.lastname: Reck-Peterson
mesh_terms: D004398:Dyneins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008954:Models, Biological; D020409:Molecular Motor Proteins / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: C075341:KIP2 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D020409:Molecular Motor Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; D004398:Dyneins; D016547:Kinesins
keywords: DNA origami; cytoskeleton; dynein; kinesin; microtubule