FAP57/WDR65 targets assembly of a subset of inner arm dyneins and connects to regulatory hubs in cilia

Jianfeng Lin, Thuc Vy Le, Katherine Augspurger, Douglas Tritschler, Raqual Bower, Gang Fu, Catherine Perrone, Eileen T. O'Toole, Kristyn VanderWaal Mills, Erin Dymek, Elizabeth Smith, Daniela Nicastro, Mary E. Porter

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Ciliary motility depends on both the precise spatial organization of multiple dynein motors within the 96 nm axonemal repeat and the highly coordinated interactions between different dyneins and regulatory complexes located at the base of the radial spokes. Mutations in genes encoding cytoplasmic assembly factors, intraflagellar transport factors, docking proteins, dynein subunits, and associated regulatory proteins can all lead to defects in dynein assembly and ciliary motility. Significant progress has been made in the identification of dynein subunits and extrinsic factors required for preassembly of dynein complexes in the cytoplasm, but less is known about the docking factors that specify the unique binding sites for the different dynein isoforms on the surface of the doublet microtubules. We have used insertional mutagenesis to identify a new locus, IDA8/BOP2, required for targeting the assembly of a subset of inner dynein arms (IDAs) to a specific location in the 96 nm repeat. IDA8 encodes flagellar-associated polypeptide (FAP)57/WDR65, a highly conserved WD repeat, coiled coil domain protein. Using high resolution proteomic and structural approaches, we find that FAP57 forms a discrete complex. Cryo-electron tomography coupled with epitope tagging and gold labeling reveal that FAP57 forms an extended structure that interconnects multiple IDAs and regulatory complexes.

Original languageEnglish (US)
Pages (from-to)2659-2680
Number of pages22
JournalMolecular biology of the cell
Volume30
Issue number21
DOIs
StatePublished - Oct 1 2019

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Dyneins
Cilia
Electron Microscope Tomography
Insertional Mutagenesis
Protein Subunits
Microtubules
Gold
Proteomics
Epitopes
Protein Isoforms
Cytoplasm
Binding Sites
Peptides
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Lin, J., Le, T. V., Augspurger, K., Tritschler, D., Bower, R., Fu, G., ... Porter, M. E. (2019). FAP57/WDR65 targets assembly of a subset of inner arm dyneins and connects to regulatory hubs in cilia. Molecular biology of the cell, 30(21), 2659-2680. https://doi.org/10.1091/mbc.E19-07-0367

FAP57/WDR65 targets assembly of a subset of inner arm dyneins and connects to regulatory hubs in cilia. / Lin, Jianfeng; Le, Thuc Vy; Augspurger, Katherine; Tritschler, Douglas; Bower, Raqual; Fu, Gang; Perrone, Catherine; O'Toole, Eileen T.; VanderWaal Mills, Kristyn; Dymek, Erin; Smith, Elizabeth; Nicastro, Daniela; Porter, Mary E.

In: Molecular biology of the cell, Vol. 30, No. 21, 01.10.2019, p. 2659-2680.

Research output: Contribution to journalArticle

Lin, J, Le, TV, Augspurger, K, Tritschler, D, Bower, R, Fu, G, Perrone, C, O'Toole, ET, VanderWaal Mills, K, Dymek, E, Smith, E, Nicastro, D & Porter, ME 2019, 'FAP57/WDR65 targets assembly of a subset of inner arm dyneins and connects to regulatory hubs in cilia', Molecular biology of the cell, vol. 30, no. 21, pp. 2659-2680. https://doi.org/10.1091/mbc.E19-07-0367
Lin, Jianfeng ; Le, Thuc Vy ; Augspurger, Katherine ; Tritschler, Douglas ; Bower, Raqual ; Fu, Gang ; Perrone, Catherine ; O'Toole, Eileen T. ; VanderWaal Mills, Kristyn ; Dymek, Erin ; Smith, Elizabeth ; Nicastro, Daniela ; Porter, Mary E. / FAP57/WDR65 targets assembly of a subset of inner arm dyneins and connects to regulatory hubs in cilia. In: Molecular biology of the cell. 2019 ; Vol. 30, No. 21. pp. 2659-2680.
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