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Cited by 5Microtubule and mitosis dynamicsCellular Mechanics and InteractionsProtein Structure and Dynamics

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Dimerization of GAS2 mediates crosslinking of microtubules and F-actin

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An, Jiancheng, Imasaki, Tsuyoshi, Narita, Akihiro, Niwa, Shinsuke, Sasaki, Ryohei, Makino, Tsukasa, Nitta, Ryo, Kikkawa, Masahide·2025·The EMBO Journal
DOI 10.1038/s44318-025-00415-2PMC PMC12084551

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Abstract

The spectraplakin family protein GAS2 was originally identified as a growth arrest-specific protein, and recent studies have revealed its involvement in multiple cellular processes. Its dual interaction with actin filaments and microtubules highlights its essential role in cytoskeletal organization, such as cell division, apoptosis, and possibly tumorigenesis. However, the structural basis of cytoskeletal dynamics regulation by GAS2 remains unclear. In this study, we present cryo-electron microscopy structures of the GAS2 type 3 calponin homology domain (CH3) in complex with F-actin at 2.8 Å resolution, thus solving the first type CH3 domain structure bound to F-actin and confirming its actin-binding activity. We also provide the first near-atomic resolution cryo-EM structure of the GAS2-GAR domain bound to microtubules and identify conserved microtubule-binding residues. Our biochemical experiments show that GAS2 promotes microtubule nucleation and polymerization, and that its C-terminal region is essential for dimerization, bundling of both F-actin and microtubules, and microtubule nucleation. As mutations leading to expression of C-terminally truncated GAS2 have been linked to hearing loss, these findings suggest that the disruption of GAS2-dependent cytoskeletal organisation could underlie auditory dysfunction.

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Metadata

Authors
An, J., Imasaki, T., Narita, A., Niwa, S., Sasaki, R., Makino, T., Nitta, R., & Kikkawa, M.
Affiliations
  • The University of Tokyo (JP)
  • Kobe University (JP)
  • Nagoya University (JP)
  • Tohoku University (JP)
Corresponding author
Kikkawa, Masahide
ORCID
Venue
The EMBO Journal
Publisher
Springer Science and Business Media LLC
Published
2025-04-01
Volume
44
Issue
10
Pages
2997-3024
ISSN
  • 0261-4189 (ISSN-L)
  • 1460-2075
DOI
10.1038/s44318-025-00415-2
PMC
PMC12084551
OpenAlex
W4409039625
Cited by
5
References
2
Citations by year
2026: 4 · 2025: 1
FWCI
2.48
Type
journal article
Language
en
Open access
Licence
Locations
  • The EMBO Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • MEXT | Japan Society for the Promotion of Science (21H04762, 21H05248, 23K27145, 21H05254, 22H05523)
  • Japan Agency for Medical Research and Development (JP22ama121002j002)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
GAS2, spectraplakins, calponin homology domain, F-actin binding, microtubule binding, protein dimerization, microtubule nucleation, microtubule bundling, actin bundling, cytoskeletal organization, cryo-EM structure, hearing loss
MeSH terms
Growth Arrest-Specific Protein 6, Actins, Animals, Humans, Microfilament Proteins, Microtubules, Models, Molecular, Protein Binding, Cryoelectron Microscopy, Intercellular Signaling Peptides and Proteins, Protein Multimerization

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