TY - JOUR
T1 - DRC2/CCDC65 is a central hub for assembly of the nexin–dynein regulatory complex and other regulators of ciliary and flagellar motility
AU - Bower, Raqual
AU - Tritschler, Douglas
AU - VanderWaal Mills, Kristyn
AU - Heuser, Thomas
AU - Nicastro, Daniela
AU - Portera, Mary E.
N1 - Funding Information:
We thank LeeAnn Higgins, Todd Markowski, and Bruce Witthun in the Center for Mass Spectrometry and Computational Proteomics at the University of Minnesota (UMN) for assistance with iTRAQ labeling, mass spectrometry, and spectral counting, and Chen Xu for training and maintenance of the Brandeis EM facility. This center is supported by multiple grants including National Science Foundation (NSF) Major Research Instrumentation grants 9871237 and NSF-DBI-0215759 as described at www.cbs.umn.edu/msp/about. We also acknowledge the University of Minnesota Supercomputing Institute for software support and data storage and Mark Sanders and the University Imaging Center for assistance with live cell imaging. We also thank Matt Laudon and the Chlamydomonas Genetics Center (UMN) for strains. Richard Linck (UMN), Mark Sanders (UMN), Ritsu Kamiya (University of Tokyo), Toshiki Yagi (Kyoto University), Pinfen Yang (Marquette University), Elizabeth Smith (Dartmouth University), and Gianni Piperno (Mount Sinai School of Medicine) generously supplied antibodies as listed in Supplemental Table S3. Preliminary reports of this work were presented at American Society for Cell Biology meetings. This work was supported by National Institutes of Health grants to M.E.P. (GM-055667) and D.N. (GM-083122).
Publisher Copyright:
© 2018 Bower et al.
PY - 2018/1/15
Y1 - 2018/1/15
N2 - The nexin–dynein regulatory complex (N-DRC) plays a central role in the regulation of ciliary and flagellar motility. In most species, the N-DRC contains at least 11 subunits, but the specific function of each subunit is unknown. Mutations in three subunits (DRC1, DRC2/CCDC65, DRC4/GAS8) have been linked to defects in ciliary motility in humans and lead to a ciliopathy known as primary ciliary dyskinesia (PCD). Here we characterize the biochemical, structural, and motility phenotypes of two mutations in the DRC2 gene of Chlamydomonas. Using high-resolution proteomic and structural approaches, we find that the C-terminal region of DRC2 is critical for the coassembly of DRC2 and DRC1 to form the base plate of N-DRC and its attachment to the outer doublet microtubule. Loss of DRC2 in drc2 mutants disrupts the assembly of several other N-DRC subunits and also destabilizes the assembly of several closely associated structures such as the inner dynein arms, the radial spokes, and the calmodulin- and spoke-associated complex. Our study provides new insights into the range of ciliary defects that can lead to PCD.
AB - The nexin–dynein regulatory complex (N-DRC) plays a central role in the regulation of ciliary and flagellar motility. In most species, the N-DRC contains at least 11 subunits, but the specific function of each subunit is unknown. Mutations in three subunits (DRC1, DRC2/CCDC65, DRC4/GAS8) have been linked to defects in ciliary motility in humans and lead to a ciliopathy known as primary ciliary dyskinesia (PCD). Here we characterize the biochemical, structural, and motility phenotypes of two mutations in the DRC2 gene of Chlamydomonas. Using high-resolution proteomic and structural approaches, we find that the C-terminal region of DRC2 is critical for the coassembly of DRC2 and DRC1 to form the base plate of N-DRC and its attachment to the outer doublet microtubule. Loss of DRC2 in drc2 mutants disrupts the assembly of several other N-DRC subunits and also destabilizes the assembly of several closely associated structures such as the inner dynein arms, the radial spokes, and the calmodulin- and spoke-associated complex. Our study provides new insights into the range of ciliary defects that can lead to PCD.
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U2 - 10.1091/mbc.E17-08-0510
DO - 10.1091/mbc.E17-08-0510
M3 - Article
C2 - 29167384
AN - SCOPUS:85041007531
SN - 1059-1524
VL - 29
SP - 137
EP - 153
JO - Molecular biology of the cell
JF - Molecular biology of the cell
IS - 2
ER -