The mitotic kinesin-14 KlpA contains a context-dependent directionality switch

Nat Commun. 2017 Jan 4:8:13999. doi: 10.1038/ncomms13999.

Abstract

Kinesin-14s are commonly known as nonprocessive minus end-directed microtubule motors that function mainly for mitotic spindle assembly. Here we show using total internal reflection fluorescence microscopy that KlpA-a kinesin-14 from Aspergillus nidulans-is a context-dependent bidirectional motor. KlpA exhibits plus end-directed processive motility on single microtubules, but reverts to canonical minus end-directed motility when anchored on the surface in microtubule-gliding experiments or interacting with a pair of microtubules in microtubule-sliding experiments. Plus end-directed processive motility of KlpA on single microtubules depends on its N-terminal nonmotor microtubule-binding tail, as KlpA without the tail is nonprocessive and minus end-directed. We suggest that the tail is a de facto directionality switch for KlpA motility: when the tail binds to the same microtubule as the motor domain, KlpA is a plus end-directed processive motor; in contrast, when the tail detaches from the microtubule to which the motor domain binds, KlpA becomes minus end-directed.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aspergillus nidulans / physiology*
  • Fungal Proteins / chemistry
  • Fungal Proteins / metabolism*
  • Kinesins / chemistry
  • Kinesins / metabolism*
  • Microscopy, Fluorescence / methods
  • Microtubules / metabolism*
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Single Molecule Imaging / methods
  • Spindle Apparatus / metabolism*
  • Time-Lapse Imaging / methods

Substances

  • Fungal Proteins
  • KLPA protein, Aspergillus nidulans
  • Recombinant Proteins
  • Kinesins