title: Measuring collective transport by defined numbers of processive and nonprocessive kinesin motors.
abstract: Intracellular transport is thought to be achieved by teams of motor proteins bound to a cargo. However, the coordination within a team remains poorly understood as a result of the experimental difficulty in controlling the number and composition of motors. Here, we developed an experimental system that links together defined numbers of motors with defined spacing on a DNA scaffold. By using this system, we linked multiple molecules of two different types of kinesin motors, processive kinesin-1 or nonprocessive Ncd (kinesin-14), in vitro. Both types of kinesins markedly increased their processivities with motor number. Remarkably, despite the poor processivity of individual Ncd motors, the coupling of two Ncd motors enables processive movement for more than 1 μm along microtubules (MTs). This improvement was further enhanced with decreasing spacing between motors. Force measurements revealed that the force generated by groups of Ncd is additive when two to four Ncd motors work together, which is much larger than that generated by single motors. By contrast, the force of multiple kinesin-1s depends only weakly on motor number. Numerical simulations and single-molecule unbinding measurements suggest that this additive nature of the force exerted by Ncd relies on fast MT binding kinetics and the large drag force of individual Ncd motors. These features would enable small groups of Ncd motors to crosslink MTs while rapidly modulating their force by forming clusters. Thus, our experimental system may provide a platform to study the collective behavior of motor proteins from the bottom up.
authors.affiliation: Advanced ICT Research Institute, National Institute of Information and Communications Technology, Kobe 651-2492, Japan. furutak@nict.go.jp
authors.forename: Ken'ya
authors.initials: K
authors.lastname: Furuta
authors.forename: Akane
authors.initials: A
authors.lastname: Furuta
authors.forename: Yoko Y
authors.initials: YY
authors.lastname: Toyoshima
authors.forename: Misako
authors.initials: M
authors.lastname: Amino
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Oiwa
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kojima
mesh_terms: D000465:Algorithms; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport / Q000502:physiology; D001703:Biophysics; D019281:Dimerization; D004926:Escherichia coli; D005453:Fluorescence; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016547:Kinesins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D056004:Molecular Dynamics Simulation; D020409:Molecular Motor Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009010:Monte Carlo Method; D015513:Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D052898:Optical Tweezers; D051381:Rats; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D020409:Molecular Motor Proteins; D015513:Oncogene Proteins; D014404:Tubulin; C502598:KIF14 protein, human; D016547:Kinesins