title: A method for multiprotein assembly in cells reveals independent action of kinesins in complex.
abstract: Teams of processive molecular motors are critical for intracellular transport and organization, yet coordination between motors remains poorly understood. Here, we develop a system using protein components to generate assemblies of defined spacing and composition inside cells. This system is applicable to studying macromolecular complexes in the context of cell signaling, motility, and intracellular trafficking. We use the system to study the emergent behavior of kinesin motors in teams. We find that two kinesin motors in complex act independently (do not help or hinder each other) and can alternate their activities. For complexes containing a slow kinesin-1 and fast kinesin-3 motor, the slow motor dominates motility in vitro but the fast motor can dominate on certain subpopulations of microtubules in cells. Both motors showed dynamic interactions with the complex, suggesting that motor-cargo linkages are sensitive to forces applied by the motors. We conclude that kinesin motors in complex act independently in a manner regulated by the microtubule track.
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Norris
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Virupakshi
authors.initials: V
authors.lastname: Soppina
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Aslan S
authors.initials: AS
authors.lastname: Dizaji
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Kristin I
authors.initials: KI
authors.lastname: Schimert
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: David
authors.initials: D
authors.lastname: Sept
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Dawen
authors.initials: D
authors.lastname: Cai
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Sivaraj
authors.initials: S
authors.lastname: Sivaramakrishnan
authors.affiliation: Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 Department of Biophysics, Department of Cell and Developmental Biology, and Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109 kjverhey@umich.edu.
authors.forename: Kristen J
authors.initials: KJ
authors.lastname: Verhey
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006801:Humans; D016547:Kinesins / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry; D015202:Protein Engineering; D055503:Protein Multimerization; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry; D051381:Rats; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D034421:Sus scrofa
chemical_list: C490575:Kif1a protein, rat; C105147:MNN4 protein, S cerevisiae; D008565:Membrane Proteins; D015800:Protozoan Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D016547:Kinesins