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Comparative Super-resolution Mapping of Basal Feet Reveals a Modular, but Distinct Architecture in Primary and Motile Cilia

Comparative Super-resolution Mapping of Basal Feet Reveals a Modular, but Distinct Architecture in Primary and Motile Cilia
Comparative Super-resolution Mapping of Basal Feet Reveals a Modular, but Distinct Architecture in Primary and Motile Cilia

In situ molecular architecture analysis of organelles and protein assemblies is essential to understanding the role of individual components and their cellular function, and to engineering new molecular functionalities. Through a super-resolution-driven approach, here we characterize the organization of the ciliary basal foot, an appendage of basal bodies whose main role is to provide a point of anchoring to the microtubule cytoskeleton. Quantitative image analysis shows that the basal foot is organized into three main regions linked by elongated coiled-coil proteins, revealing a conserved modular architecture in primary and motile cilia, but showing distinct features reflecting its specialized functions. Using domain-specific BioID proximity labeling and super-resolution imaging, we identify CEP112 as a basal foot protein and other candidate components of this assembly, aiding future investigations on the role of basal foot across different cilia systems.

airway, appendages, basal bodies, basal foot, centrosome, cilia, electron microscopy, primary ciliary dyskinesia, quantitative imaging, super-resolution imaging
1534-5807
209-223.e7
Ngueyn, Quynh
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Liu, Zhen
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Albulescu, Alexandra
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Ouyang, Hong
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Zlock, Lorna
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Coyaud, Etienne
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Laurent, Estelle
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Finkbeiner, Walter
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Moraes, Theo
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Raught, Brian
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Mennella, Vito
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Ngueyn, Quynh
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Liu, Zhen
aee11087-cc22-445d-b592-e881e70aadc8
Albulescu, Alexandra
b2b84956-2fb6-4ce3-ad99-423a6938771d
Ouyang, Hong
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Zlock, Lorna
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Coyaud, Etienne
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Laurent, Estelle
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Finkbeiner, Walter
a766f388-c3d4-4da0-bfc8-ace760569e40
Moraes, Theo
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Raught, Brian
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Mennella, Vito
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Ngueyn, Quynh, Liu, Zhen, Albulescu, Alexandra, Ouyang, Hong, Zlock, Lorna, Coyaud, Etienne, Laurent, Estelle, Finkbeiner, Walter, Moraes, Theo, Raught, Brian and Mennella, Vito (2020) Comparative Super-resolution Mapping of Basal Feet Reveals a Modular, but Distinct Architecture in Primary and Motile Cilia. Developmental Cell, 55 (2), 209-223.e7. (doi:10.1016/j.devcel.2020.09.015).

Record type: Article

Abstract

In situ molecular architecture analysis of organelles and protein assemblies is essential to understanding the role of individual components and their cellular function, and to engineering new molecular functionalities. Through a super-resolution-driven approach, here we characterize the organization of the ciliary basal foot, an appendage of basal bodies whose main role is to provide a point of anchoring to the microtubule cytoskeleton. Quantitative image analysis shows that the basal foot is organized into three main regions linked by elongated coiled-coil proteins, revealing a conserved modular architecture in primary and motile cilia, but showing distinct features reflecting its specialized functions. Using domain-specific BioID proximity labeling and super-resolution imaging, we identify CEP112 as a basal foot protein and other candidate components of this assembly, aiding future investigations on the role of basal foot across different cilia systems.

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Accepted/In Press date: 14 September 2020
e-pub ahead of print date: 9 October 2020
Published date: 26 October 2020
Additional Information: Funding Information: This project is funded by CIHR program grant #391917 to V.M.; Z.L. has been supported by the SickKids Restracomp Fellowship. The authors acknowledge volunteers for providing nasal cells for this study; Julie Avolio for help with nasal cell scraping; Jia Zhou, Cindy Fang, and Jasmine Kang assisted in data analysis; Douglas Holmyard (EM facility, the Hospital for Sick Children) prepared TEM and FIB-SEM samples and helped set up EM imaging. We thank Profs Michel Bornens, Laurence Pelletier, Iain Cheeseman, Kyung Lee, Hans-Peter Elsasser, and Tomer Avidor-Reiss laboratories for sharing antibodies and plasmids. Publisher Copyright: © 2020 Elsevier Inc.
Keywords: airway, appendages, basal bodies, basal foot, centrosome, cilia, electron microscopy, primary ciliary dyskinesia, quantitative imaging, super-resolution imaging

Identifiers

Local EPrints ID: 444603
URI: http://eprints.soton.ac.uk/id/eprint/444603
ISSN: 1534-5807
PURE UUID: 9ff6fdfe-4a71-4af3-b74a-ad7989121a2a
ORCID for Vito Mennella: ORCID iD orcid.org/0000-0002-4842-9012

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Date deposited: 27 Oct 2020 19:52
Last modified: 17 Mar 2024 05:57

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Contributors

Author: Quynh Ngueyn
Author: Zhen Liu
Author: Alexandra Albulescu
Author: Hong Ouyang
Author: Lorna Zlock
Author: Etienne Coyaud
Author: Estelle Laurent
Author: Walter Finkbeiner
Author: Theo Moraes
Author: Brian Raught
Author: Vito Mennella ORCID iD

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