Minus-end-directed motion of kinesin-coated microspheres driven by microtubule depolymerization.
Dynamic changes in microtubule (MT) length have long been thought to contribute to intracellular motility. Both the polymerization and depolymerization of tubulin have been shown to do work in vitro, but the biochemical complexity of objects moved, such as chromosomes, has complicated the identification of proteins that couple MT dynamics with motility. Work with MTs grown from and tethered to pellicles of lysed Tetrahymena has shown that disassembly-dependent movement of chromosomes in vitro can be inhibited with antibodies against the motor domain of kinesin. To study proteins that can function in disassembly-dependent motion, we have refined this motility assay, replacing chromosomes with protein-coated latex microspheres. We report here the ability of several enzymes, including kinesin, to support in vitro motility of latex microspheres on disassembling MTs (Fig. 1a). The polarity of kinesin's motor activity can be reversed by MT disassembly and interactions between a motor and a MT end can either slow or speed the rate of tubulin depolymerization.
Lombillo
V A
VA
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309.
Stewart
R J
RJ
McIntosh
J R
JR
D000818:Animals; D001704:Biopolymers; D002696:Chlamydomonas; D004023:Dictyostelium; D004330:Drosophila; D004398:Dyneins / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000502:physiology*; D008863:Microspheres; D008870:Microtubules / Q000502:physiology*; D009068:Movement; D011993:Recombinant Fusion Proteins; D013768:Tetrahymena
D001704:Biopolymers; D011993:Recombinant Fusion Proteins; D004398:Dyneins; D016547:Kinesins