α- and β-Tubulin Lattice of the Axonemal Microtubule Doublet and Binding Proteins Revealed by Single Particle Cryo-Electron Microscopy and Tomography
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Microtubule doublet (MTD) is the main skeleton of cilia/flagella. Many proteins, such as dyneins and radial spokes, bind to MTD, and generate or regulate force. While the structure of the reconstituted microtubule has been solved at atomic resolution, nature of the axonemal MTD is still unclear. There are a few hypotheses of the lattice arrangement of its α- and β-tubulins, but it has not been described how dyneins and radial spokes bind to MTD. In this study, we analyzed the three-dimensional structure of Tetrahymena MTD at ∼19 Å resolution by single particle cryo-electron microscopy. To identify α- and β-tubulins, we combined image analysis of MTD with specific kinesin decoration. This work reveals that α- and β-tubulins form a B-lattice arrangement in the entire MTD with a seam at the outer junction. We revealed the unique way in which inner arm dyneins, radial spokes, and proteins inside MTD bind and bridge protofilaments.
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The inner junction is well positioned to act as a scaffold for the radial spoke and dynein regulatory complex machinery.
“Based on near-atomic-resolution cryo-electron microscopy reconstructions of motile cilia axonemes, the inner junction is well positioned to act as a scaffold for the radial spoke and dynein regulatory complex machinery19–22.”
From The inner junction protein CFAP20 functions in motile and non-motile cilia and is critical for vision (2022) · cites this paper as [22]
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- Authors
- Maheshwari, A., Obbineni, J. M., Bui, K. H., Shibata, K., Toyoshima, Y. Y., & Ishikawa, T.
- Affiliations
- Paul Scherrer Institute (CH)
- The University of Tokyo (JP)
- Corresponding author
- Ishikawa, Takashi
- Venue
- Structure
- Publisher
- Elsevier BV
- Published
- 2015-09
- Volume
- 23
- Issue
- 9
- Pages
- 1584-1595
- ISSN
- 0969-2126 (ISSN-L)
- OpenAlex
- W2150759583
- Cited by
- 47
- References
- 42
- Citations by year
- 2026: 2 · 2025: 7 · 2024: 4 · 2023: 3 · 2022: 4 · 2021: 1 · 2020: 1 · 2019: 5 · 2018: 9 · 2017: 6 · 2016: 5
- FWCI
- 4.56
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Structure (publishedVersion)
- PubMed (publishedVersion)
- DORA PSI (Paul Scherrer Institute) (submittedVersion)
- Funders
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
- Japan Society for the Promotion of Science (25650048)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Keywords
- alpha-tubulin, beta-tubulin, radial spokes, cilia, flagella, Tetrahymena, microtubule-binding protein
- MeSH terms
- Binding Sites, Cilia, Cytoskeletal Proteins, Models, Molecular, Protein Binding, Tubulin, Protozoan Proteins, Kinesins, Tetrahymena thermophila, Crystallography, X-Ray, Cryoelectron Microscopy
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