The kinesin-5 tail domain directly modulates the mechanochemical cycle of the motor domain for anti-parallel microtubule sliding

  1. Tatyana Bodrug
  2. Elizabeth M Wilson-Kubalek
  3. Stanley Nithianantham
  4. Alex F Thompson
  5. April Alfieri
  6. Ignas Gaska
  7. Jennifer Major
  8. Garrett Debs
  9. Sayaka Inagaki
  10. Pedro Gutierrez
  11. Larisa Gheber
  12. Richard J McKenney
  13. Charles Vaughn Sindelar
  14. Ron Milligan
  15. Jason Stumpff
  16. Steven S Rosenfeld
  17. Scott T Forth
  18. Jawdat Al-Bassam  Is a corresponding author
  1. University of California, Davis, United States
  2. The Scripps Research Institute, United States
  3. University of Vermont, United States
  4. Rensselaer Polytechnic Institute, United States
  5. Lerner Research Institute, Cleveland Clinic, United States
  6. Yale University, United States
  7. Mayo Clinic, United States
  8. Ben Gurion University of the Negev, Israel

Abstract

Kinesin-5 motors organize mitotic spindles by sliding apart microtubules. They are homotetramers with dimeric motor and tail domains at both ends of a bipolar minifilament. Here, we describe a regulatory mechanism involving direct binding between tail and motor domains and its fundamental role in microtubule sliding. Kinesin-5 tails decrease microtubule-stimulated ATP-hydrolysis by specifically engaging motor domains in the nucleotide-free or ADP states. Cryo-EM reveals that tail binding stabilizes an open motor domain ATP-active site. Full-length motors undergo slow motility and cluster together along microtubules, while tail-deleted motors exhibit rapid motility without clustering. The tail is critical for motors to zipper together two microtubules by generating substantial sliding forces. The tail is essential for mitotic spindle localization, which becomes severely reduced in tail-deleted motors. Our studies suggest a revised microtubule-sliding model, in which kinesin-5 tails stabilize motor domains microtubule-bound states by slowing ATP-binding resulting in high-force production at both homotetramer ends.

Data availability

Two atomic coordinate files for Dm-KLP61F motor ATP-like MT(alpha-beta-tubulin) model is available at Protein Data Bank (PDB-ID: #XXXX. The Dm-KLP61F motor nucleotide-free MT(alpha-beta-tubulin) asymmetric unit Protein Data BankPDB-ID: #XXXXThe refined Dm-KLP61F motor AMPPNP MT cryo-EM map is available at the Electron microscopy Data bank (EMBD) EMDB ID:#XXXXand the Dm-KLP61F motor-tail nucleotide free MT cryo-EM map is available at Electron microscopy Data bank (EMBD) EMDB-iD:#XXXXDm-KLP61F motor-tail nucleotide free MT cryo-EM map (focused 3D-classification map) is available at the Electron microscopy Data bank (EMBD)EMDB-ID:#XXXX

Article and author information

Author details

  1. Tatyana Bodrug

    Department of Molecular and Cellular Biology, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Elizabeth M Wilson-Kubalek

    Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Stanley Nithianantham

    Department of Molecular and Cellular Biology, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6238-647X
  4. Alex F Thompson

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. April Alfieri

    Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Ignas Gaska

    Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Jennifer Major

    Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Garrett Debs

    Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Sayaka Inagaki

    Department of Pharmacology, Mayo Clinic, Jacksonville, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Pedro Gutierrez

    Department of Molecular and Cellular Biology, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Larisa Gheber

    Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3759-4001
  12. Richard J McKenney

    Department of Molecular and Cellular Biology, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Charles Vaughn Sindelar

    Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6646-7776
  14. Ron Milligan

    Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Jason Stumpff

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Steven S Rosenfeld

    Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Scott T Forth

    Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. Jawdat Al-Bassam

    Department of Molecular and Cellular Biology, University of California, Davis, Davis, United States
    For correspondence
    jmalbassam@ucdavis.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6625-2102

Funding

National Science Foundation (1615991)

  • Jawdat Al-Bassam

National Institutes of Health (GM110283)

  • Jawdat Al-Bassam

National Institutes of Health (GM121491)

  • Jason Stumpff

National Institutes of Health (GM130556)

  • Jason Stumpff

Israel Science Foundation (ISF 386/18)

  • Larisa Gheber

United States-Israel Binational Science Foundation (BSF-2015851)

  • Larisa Gheber

National Institutes of Health (GM130556)

  • Richard J McKenney

National Institutes of Health (GM052468)

  • Ron Milligan

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2020, Bodrug et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,856
    views
  • 505
    downloads
  • 44
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Tatyana Bodrug
  2. Elizabeth M Wilson-Kubalek
  3. Stanley Nithianantham
  4. Alex F Thompson
  5. April Alfieri
  6. Ignas Gaska
  7. Jennifer Major
  8. Garrett Debs
  9. Sayaka Inagaki
  10. Pedro Gutierrez
  11. Larisa Gheber
  12. Richard J McKenney
  13. Charles Vaughn Sindelar
  14. Ron Milligan
  15. Jason Stumpff
  16. Steven S Rosenfeld
  17. Scott T Forth
  18. Jawdat Al-Bassam
(2020)
The kinesin-5 tail domain directly modulates the mechanochemical cycle of the motor domain for anti-parallel microtubule sliding
eLife 9:e51131.
https://doi.org/10.7554/eLife.51131

Share this article

https://doi.org/10.7554/eLife.51131

Further reading

    1. Cell Biology
    2. Developmental Biology
    Pavan K Nayak, Arul Subramanian, Thomas F Schilling
    Research Article

    Mechanical forces play a critical role in tendon development and function, influencing cell behavior through mechanotransduction signaling pathways and subsequent extracellular matrix (ECM) remodeling. Here we investigate the molecular mechanisms by which tenocytes in developing zebrafish embryos respond to muscle contraction forces during the onset of swimming and cranial muscle activity. Using genome-wide bulk RNA sequencing of FAC-sorted tenocytes we identify novel tenocyte markers and genes involved in tendon mechanotransduction. Embryonic tendons show dramatic changes in expression of matrix remodeling associated 5b (mxra5b), matrilin1 (matn1), and the transcription factor kruppel-like factor 2a (klf2a), as muscles start to contract. Using embryos paralyzed either by loss of muscle contractility or neuromuscular stimulation we confirm that muscle contractile forces influence the spatial and temporal expression patterns of all three genes. Quantification of these gene expression changes across tenocytes at multiple tendon entheses and myotendinous junctions reveals that their responses depend on force intensity, duration and tissue stiffness. These force-dependent feedback mechanisms in tendons, particularly in the ECM, have important implications for improved treatments of tendon injuries and atrophy.

    1. Cell Biology
    2. Neuroscience
    Vibhavari Aysha Bansal, Jia Min Tan ... Toh Hean Ch'ng
    Research Article

    The emergence of Aβ pathology is one of the hallmarks of Alzheimer’s disease (AD), but the mechanisms and impact of Aβ in progression of the disease is unclear. The nuclear pore complex (NPC) is a multi-protein assembly in mammalian cells that regulates movement of macromolecules across the nuclear envelope; its function is shown to undergo age-dependent decline during normal aging and is also impaired in multiple neurodegenerative disorders. Yet not much is known about the impact of Aβ on NPC function in neurons. Here, we examined NPC and nucleoporin (NUP) distribution and nucleocytoplasmic transport using a mouse model of AD (AppNL-G-F/NL-G-F) that expresses Aβ in young animals. Our studies revealed that a time-dependent accumulation of intracellular Aβ corresponded with a reduction of NPCs and NUPs in the nuclear envelope which resulted in the degradation of the permeability barrier and inefficient segregation of nucleocytoplasmic proteins, and active transport. As a result of the NPC dysfunction App KI neurons become more vulnerable to inflammation-induced necroptosis – a programmed cell death pathway where the core components are activated via phosphorylation through nucleocytoplasmic shutting. Collectively, our data implicates Aβ in progressive impairment of nuclear pore function and further confirms that the protein complex is vulnerable to disruption in various neurodegenerative diseases and is a potential therapeutic target.