title: Kinesin's neck-linker determines its ability to navigate obstacles on the microtubule surface.
abstract: The neck-linker is a structurally conserved region among most members of the kinesin superfamily of molecular motor proteins that is critical for kinesin's processive transport of intracellular cargo along the microtubule surface. Variation in the neck-linker length has been shown to directly modulate processivity in different kinesin families; for example, kinesin-1, with a shorter neck-linker, is more processive than kinesin-2. Although small differences in processivity are likely obscured in vivo by the coupling of most cargo to multiple motors, longer and more flexible neck-linkers may allow different kinesins to navigate more efficiently around the many obstacles, including microtubule-associated proteins (MAPs), that are found on the microtubule surface within cells. We hypothesize that, due to its longer neck-linker, kinesin-2 can more easily navigate obstacles (e.g., MAPs) on the microtubule surface than kinesin-1. We used total internal reflection fluorescence microscopy to observe single-molecule motility from different kinesin-1 and kinesin-2 neck-linker chimeras stepping along microtubules in the absence or presence of two Tau isoforms, 3RS-Tau and 4RL-Tau, both of which are MAPs that are known to differentially affect kinesin-1 motility. Our results demonstrate that unlike kinesin-1, kinesin-2 is insensitive to the presence of either Tau isoform, and appears to have the ability to switch protofilaments while stepping along the microtubule when challenged by an obstacle, such as Tau. Thus, although kinesin-1 may be more processive, the longer neck-linker length of kinesin-2 allows it to be better optimized to navigate the complex microtubule landscape. These results provide new insight, to our knowledge, into how kinesin-1 and kinesin-2 may work together for the efficient delivery of cargo in cells.
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, Vermont.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Hoeprich
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, Vermont.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Thompson
authors.affiliation: Cell and Molecular Biology Program, University of Vermont, Burlington, Vermont.
authors.forename: Derrick P
authors.initials: DP
authors.lastname: McVicker
authors.affiliation: Department of Bioengineering, Pennsylvania State University, University Park, Pennsylvania.
authors.forename: William O
authors.initials: WO
authors.lastname: Hancock
authors.affiliation: Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, Vermont; Cell and Molecular Biology Program, University of Vermont, Burlington, Vermont. Electronic address: cberger@uvm.edu.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Berger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004331:Drosophila melanogaster; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism*; D007734:Kymography; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D017239:Paclitaxel / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D013329:Structure-Activity Relationship; D016875:tau Proteins / Q000378:metabolism
chemical_list: D016875:tau Proteins; C004805:5'-guanylylmethylenebisphosphonate; D006160:Guanosine Triphosphate; D016547:Kinesins; D017239:Paclitaxel
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