Kinesin-14 motors participate in a force balance at microtubule plus-ends to regulate dynamic instability

Allison Ogren, Sneha Parmar, Soumya R Mukherjee, Samuel Gonzalez, Melissa Plooster, Mark R McClellan, Anirudh G. Mannava, Elliott Davidson, Trisha N. Davis, Melissa K. Gardner

Research output: Contribution to journalArticlepeer-review

Abstract

Kinesin-14 molecular motors represent an essential class of proteins that bind microtubules and walk toward their minus-ends. Previous studies have described important roles for Kinesin-14 motors at microtubule minus-ends, but their role in regulating plus-end dynamics remains controversial. Kinesin-14 motors have been shown to bind the EB family of microtubule plus-end binding proteins, suggesting that these minus-end–directed motors could interact with growing microtubule plus-ends. In this work, we explored the role of minus-end–directed Kinesin-14 motor forces in controlling plus-end microtubule dynamics. In cells, a Kinesin-14 mutant with reduced affinity to EB proteins led to increased microtubule lengths. Cell-free biophysical microscopy assays were performed using Kinesin-14 motors and an EB family marker of growing microtubule plus-ends, Mal3, which revealed that when Kinesin-14 motors bound to Mal3 at growing microtubule plus-ends, the motors subsequently walked toward the minus-end, and Mal3 was pulled away from the growing microtubule tip. Strikingly, these interactions resulted in an approximately twofold decrease in the expected postinteraction microtubule lifetime. Furthermore, generic minus-end–directed tension forces, generated by tethering growing plus-ends to the coverslip using λ-DNA, led to an approximately sevenfold decrease in the expected postinteraction microtubule growth length. In contrast, the inhibition of Kinesin-14 minus-end–directed motility led to extended tip interactions and to an increase in the expected postinteraction microtubule lifetime, indicating that plus-ends were stabilized by nonmotile Kinesin-14 motors. Together, we find that Kinesin-14 motors participate in a force balance at microtubule plus-ends to regulate microtubule lengths in cells.

Original languageEnglish (US)
Article numbere2108046119
JournalProceedings of the National Academy of Sciences of the United States of America
Volume119
Issue number8
DOIs
StatePublished - Feb 22 2022

Bibliographical note

Funding Information:
supported by NIGMS Grant R01 GM040506 to T.N.D. A.O. was supported by NIH NIGMS Training Program in Muscle Research Grant T32AR007612.We thank members of the Gardner Laboratory for helpful discussions. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs and Dr. Marija Zanic for the generous gift of HSET–GFP constructs.

Funding Information:
ACKNOWLEDGMENTS. The Gardner Laboratory is supported by NIH National Institute of General Medical Sciences (NIGMS) Grant R35-GM126974. E.D. was

Publisher Copyright:
© 2022 National Academy of Sciences. All rights reserved.

Keywords

  • Dynamics
  • Kinesin
  • Microtubule
  • Motor
  • Tubulin

PubMed: MeSH publication types

  • Journal Article

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