title: Directionality of individual kinesin-5 Cin8 motors is modulated by loop 8, ionic strength and microtubule geometry.
abstract: Kinesin-5 motors fulfil essential roles in mitotic spindle morphogenesis and dynamics as slow, processive microtubule (MT) plus-end directed motors. The Saccharomyces cerevisiae kinesin-5 Cin8 was found, surprisingly, to switch directionality. Here, we have examined directionality using single-molecule fluorescence motility assays and live-cell microscopy. On spindles, Cin8 motors mostly moved slowly (∼25 nm/s) towards the midzone, but occasionally also faster (∼55 nm/s) towards the spindle poles. In vitro, individual Cin8 motors could be switched by ionic conditions from rapid (380 nm/s) and processive minus-end to slow plus-end motion on single MTs. At high ionic strength, Cin8 motors rapidly alternated directionalities between antiparallel MTs, while driving steady plus-end relative sliding. Between parallel MTs, plus-end motion was only occasionally observed. Deletion of the uniquely large insert in loop 8 of Cin8 induced bias towards minus-end motility and affected the ionic strength-dependent directional switching of Cin8 in vitro. The deletion mutant cells exhibited reduced midzone-directed motility and efficiency to support spindle elongation, indicating the importance of directionality control for the anaphase function of Cin8.
authors.affiliation: Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Adina
authors.initials: A
authors.lastname: Gerson-Gurwitz
authors.forename: Christina
authors.initials: C
authors.lastname: Thiede
authors.forename: Natalia
authors.initials: N
authors.lastname: Movshovich
authors.forename: Vladimir
authors.initials: V
authors.lastname: Fridman
authors.forename: Maria
authors.initials: M
authors.lastname: Podolskaya
authors.forename: Tsafi
authors.initials: T
authors.lastname: Danieli
authors.forename: Stefan
authors.initials: S
authors.lastname: Lakämper
authors.forename: Dieter R
authors.initials: DR
authors.lastname: Klopfenstein
authors.forename: Christoph F
authors.initials: CF
authors.lastname: Schmidt
authors.forename: Larisa
authors.initials: L
authors.lastname: Gheber
mesh_terms: D017353:Gene Deletion; D016547:Kinesins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D009068:Movement; D009994:Osmolar Concentration; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure
chemical_list: C075342:CIN8 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D016547:Kinesins
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