Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages
- Academia Sinica, Taiwan, Republic of China
- National Yang-Ming University, Taiwan, Republic of China
- Institute of Cancer Research, United Kingdom
- Memorial Sloan Kettering Cancer Center, United States
- National Taiwan University, Taiwan, Republic of China
Abstract
Subdistal appendages (sDAPs) are centriolar elements observed proximal to the distal appendages (DAPs) in vertebrates. Despite their obvious presence, structural and functional understanding of sDAPs remains elusive. Here, by combining super-resolved localization analysis and CRISPR-Cas9 genetic perturbation, we find that, although DAPs and sDAPs are primarily responsible for distinct functions in ciliogenesis and microtubule anchoring respectively, the presence of one element actually affects the positioning of the other. Specifically, we find dual layers of both ODF2 and CEP89, where their localizations are differentially regulated by DAP and sDAP integrity. DAP depletion relaxes longitudinal occupancy of sDAP protein ninein to cover the DAP region, implying a role of DAPs in sDAP positioning. Removing sDAPs alter the distal border of centrosomal γ-tubulins, illustrating a new role of sDAPs. Together, our results provide an architectural framework of sDAPs to shed light on functional understanding, surprisingly revealing the coupling between DAPs and sDAPs.
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All data generated or analysed during this study are included in the manuscript and supporting files.
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Author details
Meng-Fu Bryan Tsou
Funding
Ministry of Science and Technology, Taiwan (107-2112-M-001-037)
- Weng Man Chong
- Tzu-Yuan Chiu
- Ting-Jui Chang
- You-Pi Liu
- T Tony Yang
- Jung-Chi Liao
Ministry of Science and Technology, Taiwan (107-2313-B-001-009)
- Weng Man Chong
- Tzu-Yuan Chiu
- Ting-Jui Chang
- You-Pi Liu
- T Tony Yang
- Jung-Chi Liao
Academia Sinica (2317-1040300)
- Weng Man Chong
- Tzu-Yuan Chiu
- Ting-Jui Chang
- You-Pi Liu
- T Tony Yang
- Jung-Chi Liao
Ministry of Science and Technology, Taiwan (108-2313-B-010-001)
- Won-Jing Wang
Ministry of Science and Technology, Taiwan (108-2628-B-010-007)
- Won-Jing Wang
Ministry of Science and Technology, Taiwan (108-2638-B-010-001 -MY2)
- Won-Jing Wang
National Institutes of Health (GM088253)
- Meng-Fu Bryan Tsou
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2020, Chong et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages
eLife 9:e53580.
https://doi.org/10.7554/eLife.53580
