The inner junction complex of the cilia is an interaction hub that involves tubulin post-translational modifications
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Abstract
Microtubules are cytoskeletal structures involved in stability, transport and organization in the cell. The building blocks, the α- and β-tubulin heterodimers, form protofilaments that associate laterally into the hollow microtubule. Microtubule also exists as highly stable doublet microtubules in the cilia where stability is needed for ciliary beating and function. The doublet microtubule maintains its stability through interactions at its inner and outer junctions where its A- and B-tubules meet. Here, using cryo-electron microscopy, bioinformatics and mass spectrometry of the doublets of Chlamydomonas reinhardtii and Tetrahymena thermophila, we identified two new inner junction proteins, FAP276 and FAP106, and an inner junction-associated protein, FAP126, thus presenting the complete answer to the inner junction identity and localization. Our structural study of the doublets shows that the inner junction serves as an interaction hub that involves tubulin post-translational modifications. These interactions contribute to the stability of the doublet and hence, normal ciliary motility.
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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 [21]
The paper’s Abstract describes the inner junction as an interaction hub, and its Introduction says radial spokes and the dynein regulatory complex assemble on the doublet surface (page numbers unavailable). It does not establish that the inner junction specifically serves, or is positioned to serve, as a scaffold for those structures. (LLM, conf=0.78)
Experimental — accuracy not yet measuredProteins at or near the inner junction, including CFAP20, PACRG, CFAP45, CFAP52, CFAP90, CFAP126 and EFHC1, are positioned to form a potential scaffold for, or functionally influence, the radial spokes and dynein regulatory complex that regulates ciliary motility.
“From cryo-EM structures of Chlamydomonas and Tetrahymena motile cilia, proteins at or near the inner junction, which include CFAP20, PACRG, CFAP45, CFAP52, CFAP90, CFAP126 and EFHC119–21, are all positioned to form a potential scaffold for, or functionally influence, the radial spokes and dynein regulatory complex that regulates ciliary motility (Fig. 8).”
From The inner junction protein CFAP20 functions in motile and non-motile cilia and is critical for vision (2022) · cites this paper as [21]
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Experimental — accuracy not yet measuredThe multiple contacts of CFAP20 with ciliary axoneme tubulins and PACRG, as well as proximity to CFAP52 and other proteins at or near the inner junction, could explain the degree of conservation of CFAP20.
“The multiple contacts of CFAP20 with ciliary axoneme tubulins and PACRG, as well as proximity to CFAP52 and other proteins at or near the inner junction19–21,71 (Figs. 1b and 8), could explain this degree of conservation.”
From The inner junction protein CFAP20 functions in motile and non-motile cilia and is critical for vision (2022) · cites this paper as [21]
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Xifra‐Porxas, A., Ghosh, A., Mitsis, G. D., & Boudrias, M. (2021) · NeuroImage · doi:10.1016/j.neuroimage.2021.117822 · Cited by 59 · see citing works
Kankaanpää, A., Tolvanen, A. J., Saikkonen, P., Heikkinen, A., Laakkonen, E. K., Kaprio, J. A., Ollikainen, M., & Sillanpää, E. (2021) · The Journals of Gerontology Series A · doi:10.1093/gerona/glab337 · Cited by 61 · see citing works
Neuronal identity defines α-synuclein and tau toxicity
Praschberger, R., Kuenen, S., Schoovaerts, N., Kaempf, N., Singh, J., Janssens, J., Swerts, J., Nachman, E., Calatayud, C., Aerts, S., et al. (2023) · Neuron · doi:10.1016/j.neuron.2023.02.033 · Cited by 63 · see citing works
Tubulin Post-Translational Modifications: The Elusive Roles of Acetylation
Carmona, B., Marinho, H. S., Matos, C. L., Nolasco, S., & Soares, H. (2023) · Biology · doi:10.3390/biology12040561 · Cited by 58 · see citing works
- Authors
- Khalifa, A. A. Z., Ichikawa, M., Dai, D., Kubo, S., Black, C. S., Peri, K., McAlear, T. S., Veyron, S., Yang, S. K., Vargas, J., Bechstedt, S., Trempe, J.-F., & Bui, K. H.
- Affiliations
- McGill University (CA)
- Kyoto University (JP)
- Corresponding author
- Bui, Khanh Huy
- ORCID
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2020-01-17
- Volume
- 9
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC6994238
- OpenAlex
- W3000377151
- Cited by
- 1,676
- References
- 67
- Citations by year
- 2026: 159 · 2025: 264 · 2024: 275 · 2023: 301 · 2022: 336 · 2021: 273 · 2020: 64 · 2019: 4
- FWCI
- 142.78
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Canadian Institutes of Health Research (PJT-156354)
- Natural Sciences and Engineering Research Council of Canada (RGPIN-2016-04954, RGPIN-2018-04813, RGPIN-2017-04649)
- Canadian Institute for Advanced Research (Arzieli Global Scholar Program)
- Japan Society for the Promotion of Science
- Abdulla Al Ghurair Foundation for Education (Al Ghurair STEM program scholarship)
- Canada Research Chairs (950-229792)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
- Keywords
- tubulin post-translational modifications, Chlamydomonas reinhardtii, Tetrahymena thermophila, ciliary motility, microtubule stability, cryo-electron microscopy
- MeSH terms
- Cilia, Microtubules, Plant Proteins, Protein Processing, Post-Translational, Mass Spectrometry, Protozoan Proteins, Tetrahymena thermophila, Chlamydomonas reinhardtii, Computational Biology, Cryoelectron Microscopy
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