Intracellular functions and motile properties of bi-directional kinesin-5 Cin8 are regulated by neck linker docking

  1. Alina Goldstein-Levitin
  2. Himanshu Pandey
  3. Kanary Allhuzaeel
  4. Itamar Kass
  5. Larisa Gheber  Is a corresponding author
  1. Ben Gurion University of the Negev, Israel
  2. Monash University, Australia

Abstract

In this study, we analyzed intracellular functions and motile properties of neck-linker (NL) variants of the bi-directional S. cerevisiae kinesin-5 motor, Cin8. We also examined – by modeling – the configuration of H-bonds during NL docking. Decreasing the number of stabilizing H-bonds resulted in partially functional variants, as long as a conserved backbone H-bond at the N-latch position (proposed to stabilize the docked conformation of the NL) remained intact. Elimination of this conserved H-bond resulted in production of a non-functional Cin8 variant. Surprisingly, additional H-bond stabilization of the N-latch position, generated by replacement of the NL of Cin8 by sequences of the plus-end directed kinesin-5 Eg5, also produced a nonfunctional variant. In that variant, a single replacement of N-latch asparagine with glycine, as present in Cin8, eliminated the additional H-bond stabilization and rescued the functional defects. We conclude that exact N-latch stabilization during NL docking is critical for the function of bi-directional kinesin-5 Cin8.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. All Source data files have been provided.

Article and author information

Author details

  1. Alina Goldstein-Levitin

    Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva, Israel
    Competing interests
    No competing interests declared.
  2. Himanshu Pandey

    Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva, Israel
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0629-7525
  3. Kanary Allhuzaeel

    Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva, Israel
    Competing interests
    No competing interests declared.
  4. Itamar Kass

    Biochemistry and Molecular Biology, Monash University, Melbourne, Australia
    Competing interests
    Itamar Kass, is affiliated with InterX LTD. The author has no other competing interests to declare..
  5. Larisa Gheber

    Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva, Israel
    For correspondence
    lgheber@bgu.ac.il
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3759-4001

Funding

Israel Science Foundation (ISF-386/18)

  • Larisa Gheber

Isreal Binational Science Foundation (BSF-2015851)

  • Larisa Gheber

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

Reviewing Editor

  1. Kristen J Verhey, University of Michigan, United States

Publication history

  1. Preprint posted: May 29, 2020 (view preprint)
  2. Received: June 6, 2021
  3. Accepted: July 13, 2021
  4. Accepted Manuscript published: August 13, 2021 (version 1)
  5. Version of Record published: August 31, 2021 (version 2)

Copyright

© 2021, Goldstein-Levitin 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.

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  1. Alina Goldstein-Levitin
  2. Himanshu Pandey
  3. Kanary Allhuzaeel
  4. Itamar Kass
  5. Larisa Gheber
(2021)
Intracellular functions and motile properties of bi-directional kinesin-5 Cin8 are regulated by neck linker docking
eLife 10:e71036.
https://doi.org/10.7554/eLife.71036
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