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The Structure of Cilium Inner Junctions Revealed by Electron Cryo-tomography

View ORCID ProfileSam Li, Jose-Jesus Fernandez, Marisa D. Ruehle, Rachel A. Howard-Till, Amy Fabritius, View ORCID ProfileChad G. Pearson, David A. Agard, View ORCID ProfileMark E. Winey
doi: https://doi.org/10.1101/2024.09.09.612100
Sam Li
1Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94158, USA
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  • For correspondence: [email protected] [email protected] [email protected]
Jose-Jesus Fernandez
2Nanomaterials and Nanotechnology Research Center (CINN-CSIC), Health Research Institute of Asturias (ISPA), 33011 Oviedo, Spain
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Marisa D. Ruehle
3Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA
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Rachel A. Howard-Till
4Department of Molecular and Cellular Biology, University of California Davis, Davis, CA 95616, USA
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Amy Fabritius
4Department of Molecular and Cellular Biology, University of California Davis, Davis, CA 95616, USA
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Chad G. Pearson
3Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA
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David A. Agard
1Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94158, USA
5Chan Zuckerberg Institute for Advanced Biological Imaging, Redwood Shores, CA, USA
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Mark E. Winey
4Department of Molecular and Cellular Biology, University of California Davis, Davis, CA 95616, USA
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  • For correspondence: [email protected] [email protected] [email protected]
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Abstract

The cilium is a microtubule-based organelle critical for many cellular functions. Its assembly initiates at a basal body and continues as an axoneme that projects out of the cell to form a functional cilium. This assembly process is tightly regulated. However, our knowledge of the molecular architecture and the mechanism of assembly is limited. By applying electron cryo-tomography and subtomogram averaging, we obtained subnanometer resolution structures of the inner junction in three distinct regions of the cilium: the proximal region of the basal body, the central core of the basal body, and the flagellar axoneme. The structures allowed us to identify several basal body and axoneme components. While a few proteins are distributed throughout the entire length of the organelle, many are restricted to particular regions of the cilium, forming intricate local interaction networks and bolstering local structural stability. Finally, by knocking out a critical basal body inner junction component Poc1, we found the triplet MT was destabilized, resulting in a defective structure. Surprisingly, several axoneme-specific components were found to “infiltrate” into the mutant basal body. Our findings provide molecular insight into cilium assembly at its inner junctions, underscoring its precise spatial regulation.

Competing Interest Statement

The authors have declared no competing interest.

  • Abbreviations

    BB
    basal body
    TMT
    triplet microtubule
    DMT
    doublet microtubule
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    Posted September 09, 2024.
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    The Structure of Cilium Inner Junctions Revealed by Electron Cryo-tomography
    Sam Li, Jose-Jesus Fernandez, Marisa D. Ruehle, Rachel A. Howard-Till, Amy Fabritius, Chad G. Pearson, David A. Agard, Mark E. Winey
    bioRxiv 2024.09.09.612100; doi: https://doi.org/10.1101/2024.09.09.612100
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    The Structure of Cilium Inner Junctions Revealed by Electron Cryo-tomography
    Sam Li, Jose-Jesus Fernandez, Marisa D. Ruehle, Rachel A. Howard-Till, Amy Fabritius, Chad G. Pearson, David A. Agard, Mark E. Winey
    bioRxiv 2024.09.09.612100; doi: https://doi.org/10.1101/2024.09.09.612100

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