Diffusive tail anchorage determines velocity and force produced by kinesin-14 between crosslinked microtubules

Annemarie Lüdecke; Anja-Maria Seidel; Marcus Braun; Zdenek Lansky; Stefan Diez

doi:10.1038/s41467-018-04656-0

Diffusive tail anchorage determines velocity and force produced by kinesin-14 between crosslinked microtubules

Nat Commun. 2018 Jun 7;9(1):2214. doi: 10.1038/s41467-018-04656-0.

Authors

Annemarie Lüdecke¹, Anja-Maria Seidel¹, Marcus Braun^{2

3}, Zdenek Lansky^{4

5}, Stefan Diez^{6

7}

Affiliations

¹ B CUBE-Center for Molecular Bioengineering, Technische Universität Dresden, Arnoldstr. 18, 01307, Dresden, Germany.
² B CUBE-Center for Molecular Bioengineering, Technische Universität Dresden, Arnoldstr. 18, 01307, Dresden, Germany. marcus.braun@ibt.cas.cz.
³ Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50, Vestec, Czech Republic. marcus.braun@ibt.cas.cz.
⁴ B CUBE-Center for Molecular Bioengineering, Technische Universität Dresden, Arnoldstr. 18, 01307, Dresden, Germany. zdenek.lansky@ibt.cas.cz.
⁵ Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50, Vestec, Czech Republic. zdenek.lansky@ibt.cas.cz.
⁶ B CUBE-Center for Molecular Bioengineering, Technische Universität Dresden, Arnoldstr. 18, 01307, Dresden, Germany. stefan.diez@tu-dresden.de.
⁷ Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307, Dresden, Germany. stefan.diez@tu-dresden.de.

Abstract

Form and function of the mitotic spindle depend on motor proteins that crosslink microtubules and move them relative to each other. Among these are kinesin-14s, such as Ncd, which interact with one microtubule via their non-processive motor domains and with another via their diffusive tail domains, the latter allowing the protein to slip along the microtubule surface. Little is known about the influence of the tail domains on the protein's performance. Here, we show that diffusive anchorage of Ncd's tail domains impacts velocity and force considerably. Tail domain slippage reduced velocities from 270 nm s^-1 to 60 nm s^-1 and forces from several piconewtons to the sub-piconewton range. These findings challenge the notion that kinesin-14 may act as an antagonizer of other crosslinking motors, such as kinesin-5, during mitosis. It rather suggests a role of kinesin-14 as a flexible element, pliantly sliding and crosslinking microtubules to facilitate remodeling of the mitotic spindle.

Publication types

Research Support, Non-U.S. Gov't
Video-Audio Media

MeSH terms

Drosophila Proteins / isolation & purification
Drosophila Proteins / metabolism*
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / isolation & purification
Green Fluorescent Proteins / metabolism
Kinesins / isolation & purification
Kinesins / metabolism*
Microtubule-Associated Proteins / genetics
Microtubule-Associated Proteins / isolation & purification
Microtubule-Associated Proteins / metabolism
Microtubules / metabolism*
Mitosis / physiology*
Optical Tweezers
Protein Binding / physiology
Protein Domains
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / isolation & purification
Saccharomyces cerevisiae Proteins / metabolism
Spindle Apparatus / metabolism

Substances

Ase1 protein, S cerevisiae
Drosophila Proteins
Microtubule-Associated Proteins
Recombinant Proteins
Saccharomyces cerevisiae Proteins
ncd protein, Drosophila
Green Fluorescent Proteins
Kinesins