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- W2912140686 abstract "The subdistal appendages (SDAs) are centriolar structures important to microtubule (MT) nucleation and anchoring. Recent studies suggest its role in linking submerged primary cilia to Golgi. Despite its importance, little is known about their assembly geometry and structure-function relationship. Here, we use dSTORM superresolution microscopy to resolve the organization of multiple SDA proteins and centriole-associated proteins in mammalian cells. dSTORM imaging reveals that ODF2, a core SDA protein, localizes closed to the centriole wall while Cep128 and centriolin form ring-like structure building upon it. CCDC68, Cep170 and Ninein are two layered structures with one layer in the SDA region (the SDA layer) surrounding the core SDA proteins and the other layer residing close to the centriole proximal end (the proximal layer). Dual-color dSTORM shows that these SDA proteins form a triangular structure rooted from the centriole wall, mapping to the SDA architecture resolved with electron microscopy. Interestingly, dSTORM imaging of γ-tubulin, the key MT nucleating element, is found to be enriched between the SDA and the proximal layer of Cep170 at the location where MT fibers are observed. Besides, MT fibers are also found to attach to various centriole elements including the DAs, SDAs and the proximal end of the centriole. Depletion of SDAs upon Cep128 knockout suppresses MT attachment to SDAs but not DAs. dSTORM imaging also reveals that AKAP450, a scaffold protein connecting the centrosome and Golgi, forms a cup like structure surrounding the entire centriole via its interaction with Cep170. Upon Cep170 depletion, the centrosomal AKAP450 is abolished, causing detachment of the mother centriole from Golgi. Together, using dSTORM, we systematically disclosed the structural organization of SDAs and illustrated the structural function of SDAs in MT anchoring and centriole-Golgi association." @default.
- W2912140686 created "2019-02-21" @default.
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- W2912140686 date "2019-02-01" @default.
- W2912140686 modified "2023-09-27" @default.
- W2912140686 title "Super-Resolution Microscopy Reveals the Molecular Architecture of Centriole Subdistal Appendages and Its Role in Microtubule/Golgi Anchoring" @default.
- W2912140686 doi "https://doi.org/10.1016/j.bpj.2018.11.737" @default.
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