Structural Basis of Backwards Motion in Kinesin-1-Kinesin-14 Chimera: Implication for Kinesin-14 Motility

Masahiko Yamagishi; Hideki Shigematsu; Takeshi Yokoyama; Masahide Kikkawa; Mitsuhiro Sugawa; Mari Aoki; Mikako Shirouzu; Junichiro Yajima; Ryo Nitta

doi:10.1016/j.str.2016.05.021

Structural Basis of Backwards Motion in Kinesin-1-Kinesin-14 Chimera: Implication for Kinesin-14 Motility

Structure. 2016 Aug 2;24(8):1322-1334. doi: 10.1016/j.str.2016.05.021. Epub 2016 Jul 21.

Authors

Masahiko Yamagishi¹, Hideki Shigematsu², Takeshi Yokoyama², Masahide Kikkawa³, Mitsuhiro Sugawa¹, Mari Aoki², Mikako Shirouzu², Junichiro Yajima⁴, Ryo Nitta⁵

Affiliations

¹ Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan.
² RIKEN Center for Life Science Technologies, Tsurumi-ku, Yokohama 230-0045, Japan.
³ Department of Cell Biology and Anatomy, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; CREST, JST, Bunkyo-ku, Tokyo 113-0033, Japan.
⁴ Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan. Electronic address: [email protected].
⁵ RIKEN Center for Life Science Technologies, Tsurumi-ku, Yokohama 230-0045, Japan. Electronic address: [email protected].

PMID: 27452403
DOI: 10.1016/j.str.2016.05.021

Abstract

Kinesin-14 is a unique minus-end-directed microtubule-based motor. A swinging motion of a class-specific N-terminal neck helix has been proposed to produce minus-end directionality. However, it is unclear how swinging of the neck helix is driven by ATP hydrolysis utilizing the highly conserved catalytic core among all kinesins. Here, using a motility assay, we show that in addition to the neck helix, the conserved five residues at the C-terminal region in kinesin-14, namely the neck mimic, are necessary to give kinesin-1 an ability to reverse its directionality toward the minus end of microtubules. Our structural analyses further demonstrate that the C-terminal neck mimic, in cooperation with conformational changes in the catalytic core during ATP binding, forms a kinesin-14 bundle with the N-terminal neck helix to swing toward the minus end of microtubules. Thus, the neck mimic plays a crucial role in coupling the chemical ATPase reaction with the mechanical cycle to produce the minus-end-directed motility of kinesin-14.

MeSH terms

Adenosine Diphosphate / chemistry*
Adenosine Diphosphate / metabolism
Adenosine Triphosphate / chemistry*
Adenosine Triphosphate / metabolism
Amino Acid Sequence
Animals
Catalytic Domain
Crystallography, X-Ray
Drosophila Proteins / chemistry*
Drosophila Proteins / genetics
Drosophila Proteins / metabolism
Drosophila melanogaster / chemistry
Drosophila melanogaster / metabolism
Gene Expression
Kinesins / chemistry*
Kinesins / genetics
Kinesins / metabolism
Microtubules / metabolism*
Microtubules / ultrastructure
Molecular Docking Simulation
Molecular Dynamics Simulation
Motion
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Protein Isoforms / chemistry
Protein Isoforms / genetics
Protein Isoforms / metabolism
Rats
Recombinant Fusion Proteins / chemistry*
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Thermodynamics

Substances

Drosophila Proteins
Protein Isoforms
Recombinant Fusion Proteins
ncd protein, Drosophila
Adenosine Diphosphate
Adenosine Triphosphate
KIF5C protein, rat
Kinesins