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

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Dimerization GAS2 mediates microtubule and F-actin crosslinking

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An, Jiancheng, Imasaki, Tsuyoshi, Narita, Akihiro, Niwa, Shinsuke, Sasaki, Ryohei, Makino, Tsukasa, Nitta, Ryo, Kikkawa, Masahide·2024·The EMBO Journal
DOI 10.1101/2024.08.19.608523PMC PMC12084551

Preprint — published as 10.1038/s44318-025-00415-2 · Paper page

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Abstract

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 bases by which GAS2 regulates cytoskeletal dynamics remain unclear. In this study, we present cryo-EM structures of the GAS2- CH3 domain in complex with F-actin at 2.8 Å resolution, representing the first type 3 CH domain structure bound to F-actin, 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 identified conserved microtubule-binding residues. Our biochemical experiments show that GAS2 promotes microtubule nucleation and polymerization and its C-terminal region is essential for dimerization, bundling of both F-actin and microtubules, and microtubule nucleation. Based on these results, we propose how GAS2 controls cytoskeletal organization.

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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
openRxiv
Published
2024-08-19
Volume
44
Issue
10
Pages
2997-3024
ISSN
0261-4189 (ISSN-L)
DOI
10.1101/2024.08.19.608523
PMC
PMC12084551
OpenAlex
W4409039625
Cited by
5
References
1
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
  • Japan Agency for Medical Research and Development (JP22ama121002j002)
  • Ministry of Education, Culture, Sports, Science and Technology (22H05523, 23K27145, 21H05254, 21H05248, 21H04762)
  • Japan Society for the Promotion of Science (21H05248, 21H05254, 21H05247, 22K06809, 22H05523, 23K06315, 23K27145, 21H04762)
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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