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Mol Biol Cell. 2019 Sep 4:mbcE19070367. doi: 10.1091/mbc.E19-07-0367. [Epub ahead of print]

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

Author information

1
Departments of Cell Biology and Biophysics, University of Texas Southwestern Medical Center, 6000 Harry Hines Blvd, Dallas, TX 75390.
2
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455.
3
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309.
4
Department of Biological Sciences, Class of 1978 Life Sciences Center Dartmouth College, Hanover NH 03755.

Abstract

Ciliary motility depends on both the precise spatial organization of multiple dynein motors within the 96 nm axonemal repeat, and 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 pre-assembly 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 to a specific location in the 96 nm repeat. IDA8 encodes FAP57/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 inner dynein arms and regulatory complexes. Video S1 Video S1 Video of a wild-type cell swimming forward with an asymmetric waveform. Video S2 Video S2 Video of an ida8-1 cell swimming forward with an asymmetric waveform. Video S3 Video S3 Video of a bop2-1 cell swimming forward with an asymmetric waveform. Video S4 Video S4 Video of an ida8-1; Fap57-HA rescued cell swimming forward with an asymmetric waveform. Video S5 Video S5 Video of a bop2-1, FAP57-HA rescued cell swimming forward with an asymmetric waveform. Video S6 Video S6 Video of a pf10 cell swimming with an abnormal waveform. Video S7 Video S7 Video of an ida8-1; pf10 cell swimming with a variable waveform. Video S8 Video S8 Video of a bop2-1; pf10 cell swimming with a variable waveform.

PMID:
31483737
DOI:
10.1091/mbc.E19-07-0367

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