ArticleThe Journal of biological chemistry2015
In situ localization of N and C termini of subunits of the flagellar nexin-dynein regulatory complex (N-DRC) using SNAP tag and cryo-electron tomography.
Article in The Journal of biological chemistry, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 99 citations in OpenAlex.
- The New Nexin-Dynein Regulatory Complex Component CCDC153 Is Dispensable for Ciliary Motility and Fertility in Mice.Cytoskeleton (Hoboken, N.J.) · 2026Article
- Molecular tags for electron cryo-tomography.Emerging topics in life sciences · 2025Review
- Advances in high-resolution imaging of centrioles and cilia.Journal of cell science · 2025Review
- Advances in Imaging Techniques for Mammalian/Human Ciliated Cell's Cilia: Insights into Structure, Function, and Dynamics.Biology · 2025Review
- Assembly of FAP93 at the proximal axoneme in Chlamydomonas cilia.Cytoskeleton (Hoboken, N.J.) · 2024Article
- Integrative analysis of bulk and single-cell RNA sequencing reveals the gene expression profile and the critical signaling pathways of type II CPAM.Cell & bioscience · 2024Article
- The MBO2/FAP58 heterodimer stabilizes assembly of inner arm dyneinMolecular biology of the cell · 2024Article
- The underlying molecular mechanism of ciliated epithelium dysfunction and TGF-β signaling in children with congenital pulmonary airway malformations.Scientific reports · 2024Article
- Review
- Integrated modeling of the Nexin-dynein regulatory complex reveals its regulatory mechanism.Nature communications · 2023Article
- Integrated modeling of the Nexin-dynein regulatory complex reveals its regulatory mechanism.bioRxiv : the preprint server for biology · 2023Article
- Cryo-electron tomography on focused ion beam lamellae transforms structural cell biology.Nature methods · 2023Review
- In vivo imaging reveals independent intraflagellar transport of the nexin-dynein regulatory complex subunits DRC2 and DRC4.Molecular biology of the cell · 2023Article
- Electron microscopy for imaging organelles in plants and algae.Plant physiology · 2022Article
- Structural organization of the C1b projection within the ciliary central apparatus.Journal of cell science · 2021Article
- Structural organization of the intermediate and light chain complex of Chlamydomonas ciliary I1 dynein.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2021Article
- Structural insights into the architecture and assembly of eukaryotic flagella.Microbial cell (Graz, Austria) · 2020Review
- In situ structure determination at nanometer resolution using TYGRESS.Nature methods · 2020Article
- Rare Human Diseases: Model Organisms in Deciphering the Molecular Basis of Primary Ciliary Dyskinesia.Cells · 2019Review
- Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus.The Journal of cell biology · 2019Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
Cryo-electron tomography (cryo-ET) has reached nanoscale resolution for in situ three-dimensional imaging of macromolecular complexes and organelles. Yet its current resolution is not sufficient to precisely localize or identify most proteins in situ; for example, the location and arrangement of components of the nexin-dynein regulatory complex (N-DRC), a key regulator of ciliary/flagellar motility that is conserved from algae to humans, have remained elusive despite many cryo-ET studies of cilia and flagella. Here, we developed an in situ localization method that combines cryo-ET/subtomogram averaging with the clonable SNAP tag, a widely used cell biological probe to visualize fusion proteins by fluorescence microscopy. Using this hybrid approach, we precisely determined the locations of the N and C termini of DRC3 and the C terminus of DRC4 within the three-dimensional structure of the N-DRC in Chlamydomonas flagella. Our data demonstrate that fusion of SNAP with target proteins allowed for protein localization with high efficiency and fidelity using SNAP-linked gold nanoparticles, without disrupting the native assembly, structure, or function of the flagella. After cryo-ET and subtomogram averaging, we localized DRC3 to the L1 projection of the nexin linker, which interacts directly with a dynein motor, whereas DRC4 was observed to stretch along the N-DRC base plate to the nexin linker. Application of the technique developed here to the N-DRC revealed new insights into the organization and regulatory mechanism of this complex, and provides a valuable tool for the structural dissection of macromolecular complexes in situ.
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