TY - JOUR
T1 - Structural insights into the cause of human RSPH4A primary ciliary dyskinesia
AU - Zhao, Yanhe
AU - Pinskey, Justine
AU - Lin, Jianfeng
AU - Yin, Weining
AU - Sears, Patrick R.
AU - Daniels, Leigh A.
AU - Zariwala, Maimoona A.
AU - Knowles, Michael R.
AU - Ostrowski, Lawrence E.
AU - Nicastro, Daniela
N1 - Funding Information:
We thank Chen Xu for providing EM training and management of the electron microscopes in the Louise Mashal Gabbay Cellular Visualization Facility at Brandeis University. We thank David Mastronarde and John Heumann (University of Colorado at Boulder) for continued development of image processing tools, including PEET classification. We also thank the University of North Carolina Cystic Fibrosis Cell and Tissue Culture Core (supported by R026-CR11 from the Cystic Fibrosis Foundation and NIH DK065988) for providing samples and expertise. This research benefitted from the computational resources provided by the BioHPC supercomputing facility located in the Lyda Hill Department of Bioinformatics at UT Southwestern Medical Center. This work was supported by funding from the National Institutes of Health (GM083122 to D.N., 2R01HL117836 to L.E.O. and M.A.Z., 2U54HL096458 and 5R01HL071798 to M.R.K. and M.A.Z., and F32GM137470 to J.P.). The Genetic Disorders of Mucociliary Clearance Consortium (U54HL096458) is part of the National Center for Advancing Translational Sciences (NCATS) Rare Diseases Clinical Research Network (RDCRN) and is supported by the RDCRN Data Management and Coordinating Center (DMCC) (U2CTR002818). RDCRN is an initiative of the Office of Rare Diseases Research (ORDR) funded through a collaboration between NCATS and the National Heart, Lung, and Blood Institute (NHLBI). Additional support was provided by the Cancer Prevention and Research Institute of Texas Grant RR140082 (D.N), by the March of Dimes Foundation (to D.N.), and by a private donation from Arnold Cohen (to D.N.). Cryo-ET data for the RSPH1 and RSPH4A patient samples have been deposited with the EM Data Bank under accession codes EMD-22874 and EMD-22875, respectively.
Publisher Copyright:
© 2021 Zhao et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution-Noncommercial-Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
PY - 2021/5/27
Y1 - 2021/5/27
N2 - Cilia and flagella are evolutionarily conserved eukaryotic organelles involved in cell motility and signaling. In humans, mutations in Radial Spoke Head Component 4A (RSPH4A) can lead to primary ciliary dyskinesia (PCD), a life-shortening disease characterized by chronic respiratory tract infections, abnormal organ positioning, and infertility. Despite its importance for human health, the location of RSPH4A in human cilia has not been resolved, and the structural basis of RSPH4A-/- PCD remains elusive. Here, we present the native three-dimensional structure of RSPH4A-/- human respiratory cilia using samples collected noninvasively from a PCD patient. Using cryo-electron tomography (cryo-ET) and subtomogram averaging, we compared the structures of control and RSPH4A-/- cilia, revealing primary defects in two of the three radial spokes (RSs) within the axonemal repeat and secondary (heterogeneous) defects in the central pair complex. Similar to RSPH1-/- cilia, the radial spoke heads of RS1 and RS2, but not RS3, were missing in RSPH4A-/- cilia. However, RSPH4A-/- cilia also exhibited defects within the arch domains adjacent to the RS1 and RS2 heads, which were not observed with RSPH1 loss. Our results provide insight into the underlying structural basis for RSPH4A-/- PCD and highlight the benefits of applying cryo-ET directly to patient samples for molecular structure determination.
AB - Cilia and flagella are evolutionarily conserved eukaryotic organelles involved in cell motility and signaling. In humans, mutations in Radial Spoke Head Component 4A (RSPH4A) can lead to primary ciliary dyskinesia (PCD), a life-shortening disease characterized by chronic respiratory tract infections, abnormal organ positioning, and infertility. Despite its importance for human health, the location of RSPH4A in human cilia has not been resolved, and the structural basis of RSPH4A-/- PCD remains elusive. Here, we present the native three-dimensional structure of RSPH4A-/- human respiratory cilia using samples collected noninvasively from a PCD patient. Using cryo-electron tomography (cryo-ET) and subtomogram averaging, we compared the structures of control and RSPH4A-/- cilia, revealing primary defects in two of the three radial spokes (RSs) within the axonemal repeat and secondary (heterogeneous) defects in the central pair complex. Similar to RSPH1-/- cilia, the radial spoke heads of RS1 and RS2, but not RS3, were missing in RSPH4A-/- cilia. However, RSPH4A-/- cilia also exhibited defects within the arch domains adjacent to the RS1 and RS2 heads, which were not observed with RSPH1 loss. Our results provide insight into the underlying structural basis for RSPH4A-/- PCD and highlight the benefits of applying cryo-ET directly to patient samples for molecular structure determination.
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U2 - 10.1091/MBC.E20-12-0806
DO - 10.1091/MBC.E20-12-0806
M3 - Article
C2 - 33852348
AN - SCOPUS:85107163757
SN - 1059-1524
VL - 32
SP - 1202
EP - 1209
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
IS - 12
ER -