title: Ensemble and single-molecule dynamics of IFT dynein in Caenorhabditis elegans cilia.
abstract: Cytoplasmic dyneins drive microtubule-based, minus-end directed transport in eukaryotic cells. Whereas cytoplasmic dynein 1 has been widely studied, IFT dynein has received far less attention. Here, we use fluorescence microscopy of labelled motors in living Caenorhabditis elegans to investigate IFT-dynein motility at the ensemble and single-molecule level. We find that while the kinesin composition of motor ensembles varies along the track, the amount of dynein remains relatively constant. Remarkably, this does not result in directionality changes of cargo along the track, as has been reported for other opposite-polarity, tug-of-war motility systems. At the single-molecule level, IFT-dynein trajectories reveal unexpected dynamics, including diffusion at the base, and pausing and directional switches along the cilium. Stochastic simulations show that the ensemble IFT-dynein distribution depends upon the probability of single-motor directional switches. Our results provide quantitative insight into IFT-dynein dynamics in vivo, shedding light on the complex functioning of dynein motors in general.
authors.affiliation: Department of Physics and LaserLaB Amsterdam, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam 1081 HV, The Netherlands.
authors.forename: Jona
authors.initials: J
authors.lastname: Mijalkovic
authors.affiliation: Department of Physics and LaserLaB Amsterdam, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam 1081 HV, The Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Prevo
authors.affiliation: Department of Physics and LaserLaB Amsterdam, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam 1081 HV, The Netherlands.
authors.forename: Felix
authors.initials: F
authors.lastname: Oswald
authors.affiliation: Department of Physics and LaserLaB Amsterdam, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam 1081 HV, The Netherlands.
authors.forename: Pierre
authors.initials: P
authors.lastname: Mangeol
authors.affiliation: Department of Physics and LaserLaB Amsterdam, Vrije Universiteit Amsterdam, De Boelelaan 1081, Amsterdam 1081 HV, The Netherlands.
authors.forename: Erwin J G
authors.initials: EJ
authors.lastname: Peterman
mesh_terms: D000818:Animals; D001692:Biological Transport; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D002923:Cilia / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism; D008954:Models, Biological; D000072760:Single Molecule Imaging; D013269:Stochastic Processes
chemical_list: D029742:Caenorhabditis elegans Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004398:Dyneins; D016547:Kinesins