Kinesin-1 walks along microtubules by alternating ATP hydrolysis and movement of its two motor domains (“head”). The detached head preferentially binds to the forward tubulin-binding site after ATP binds to the microtubule-bound head, but the mechanism preventing premature microtubule binding while the partner head awaits ATP remains unknown. Here, we examined the role of the neck linker, the segment connecting two heads, in this mechanism. Structural analyses of the nucleotide-free head revealed a bulge just ahead of the neck linker’s base, creating an asymmetric constraint on its mobility. While the neck linker can stretch freely backward, it must navigate around this bulge to extend forward. We hypothesized that increased neck linker tension suppresses premature binding of the tethered head, which was supported by molecular dynamics simulations and single-molecule fluorescence assays. These findings demonstrate a tension-dependent allosteric mechanism that coordinates the movement of two heads, where neck linker tension modulates the allosteric conformational changes rather than directly affecting the nucleotide state.
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April 29 2025
Tension-induced suppression of allosteric conformational changes coordinates kinesin-1 stepping
Tsukasa Makino
,
Tsukasa Makino
(Conceptualization, Formal analysis, Investigation, Methodology, Visualization)
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
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Ryo Kanada
,
Ryo Kanada
(Formal analysis, Investigation, Visualization, Writing - original draft)
4Department of Biophysics,
Graduate School of Science, Kyoto University
, Kyoto, Japan
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Teppei Mori
,
Teppei Mori
(Data curation, Formal analysis, Investigation, Resources, Software, Visualization)
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
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Ken-ichi Miyazono
,
Ken-ichi Miyazono
(Investigation)
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
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Yuta Komori
,
Yuta Komori
(Data curation, Formal analysis, Methodology, Software)
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
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Haruaki Yanagisawa
,
Haruaki Yanagisawa
(Formal analysis, Investigation, Methodology, Software)
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
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Shoji Takada
,
Shoji Takada
(Supervision, Writing - review & editing)
4Department of Biophysics,
Graduate School of Science, Kyoto University
, Kyoto, Japan
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Masaru Tanokura
,
Masaru Tanokura
(Conceptualization, Funding acquisition, Project administration, Resources, Software, Supervision, Validation, Writing - original draft, Writing - review & editing)
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
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Masahide Kikkawa
,
Masahide Kikkawa
(Conceptualization, Supervision, Writing - original draft, Writing - review & editing)
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
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Michio Tomishige
(Conceptualization, Funding acquisition, Methodology, Project administration, Supervision, Validation, Visualization, Writing - original draft, Writing - review & editing)
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
5Department of Physical Sciences,
College of Science and Engineering, Aoyama Gakuin University
, Sagamihara, Japan
Correspondence to Michio Tomishige: [email protected]
Search for other works by this author on:
Tsukasa Makino
Conceptualization, Formal analysis, Investigation, Methodology, Visualization
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
Ryo Kanada
Formal analysis, Investigation, Visualization, Writing - original draft
4Department of Biophysics,
Graduate School of Science, Kyoto University
, Kyoto, Japan
Teppei Mori
Data curation, Formal analysis, Investigation, Resources, Software, Visualization
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
Ken-ichi Miyazono
Investigation
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
Yuta Komori
Data curation, Formal analysis, Methodology, Software
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
Haruaki Yanagisawa
Formal analysis, Investigation, Methodology, Software
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
Shoji Takada
Supervision, Writing - review & editing
4Department of Biophysics,
Graduate School of Science, Kyoto University
, Kyoto, Japan
Masaru Tanokura
Conceptualization, Funding acquisition, Project administration, Resources, Software, Supervision, Validation, Writing - original draft, Writing - review & editing
2Department of Applied Biological Chemistry,
Graduate School of Agricultural and Life Sciences, The University of Tokyo
, Tokyo, Japan
Masahide Kikkawa
Conceptualization, Supervision, Writing - original draft, Writing - review & editing
3Department of Cell Biology and Anatomy,
Graduate School of Medicine, The University of Tokyo
, Tokyo, Japan
Michio Tomishige
Conceptualization, Funding acquisition, Methodology, Project administration, Supervision, Validation, Visualization, Writing - original draft, Writing - review & editing
1Department of Applied Physics,
The University of Tokyo
, Tokyo, Japan
5Department of Physical Sciences,
College of Science and Engineering, Aoyama Gakuin University
, Sagamihara, Japan
Correspondence to Michio Tomishige: [email protected]
Disclosures: The authors declare no competing interests exist.
Received:
January 30 2025
Revision Received:
March 18 2025
Accepted:
April 08 2025
Online ISSN: 1540-8140
Print ISSN: 0021-9525
Funding
Funder(s):
Human Frontier Science Program
Funder(s):
Ministry of Education, Culture, Sports, Science and Technology
Funder(s):
Asahi Glass Foundation
Funder(s):
National Project on Protein Structural and Functional Analyses
© 2025 Makino et al.
2025
Makino et al.
This article is distributed under the terms as described at https://rupress.org/pages/terms102024/.
J Cell Biol (2025) 224 (7): e202501253.
Article history
Received:
January 30 2025
Revision Received:
March 18 2025
Accepted:
April 08 2025
Citation
Tsukasa Makino, Ryo Kanada, Teppei Mori, Ken-ichi Miyazono, Yuta Komori, Haruaki Yanagisawa, Shoji Takada, Masaru Tanokura, Masahide Kikkawa, Michio Tomishige; Tension-induced suppression of allosteric conformational changes coordinates kinesin-1 stepping. J Cell Biol 7 July 2025; 224 (7): e202501253. doi: https://doi.org/10.1083/jcb.202501253
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