De novo identification of mammalian ciliary motility proteins using cryo-EM

Accepted/In Press date: 7 October 2021

Published date: 11 November 2021

Additional Information: Funding Information: Cryo-EM data were collected at The Harvard Cryo-EM Center for Structural Biology. We thank Sarah Sterling, Richard Walsh, and Shaun Rawson for microscopy assistance; Ross Tomaino (Harvard Medical School) for mass spectrometry analysis; Y.L. Chong (Institute of Molecular and Cell Biology) for identification of zebrafish mutants; the A ∗ STAR Microscopy Platform and the National University of Singapore EM unit for TEM analysis of zebrafish KV; staff at the Mary Lyon Centre for mouse husbandry, genotyping, and histology; Henrik Westerberg and Jenni Vibert (MRC Harwell Institute) for help with particle image velocimetry; Claire Jackson, Janice Coles, Liz Adams, and Peter Lackie (PCD Centre, University of Southampton) for assessing mouse tracheal ciliary beating and TEM images; Elspeth Bruford (HGNC) for gene nomenclature discussions; and Marie Bao for comments. P.A. and D.W.M. were supported by University of Sheffield-A ∗ STAR Singapore and University of Manchester-A ∗ STAR Singapore doctoral studentships, respectively. This work was supported by awards from the UK Medical Research Council ( MC_U142670370 to D.P.N.), A ∗ STAR Singapore ( SC15-R0010 to S.R.), National Institutes of Health ( NIGMS 1R01GM141109 ), the LouLou Foundation , the E. Matilda Ziegler Foundation for the Blind , the Smith Family Foundation , and the Pew Charitable Trusts (to A.B.). The National PCD Centre in Southampton is commissioned and funded by NHS England ; PCD research is supported by the National Institute for Health Research ( RfPB PB-PG-1215-20014 and 200470 ) and The AAIR Charity (Reg. No. 1129698 ). Funding Information: Cryo-EM data were collected at The Harvard Cryo-EM Center for Structural Biology. We thank Sarah Sterling, Richard Walsh, and Shaun Rawson for microscopy assistance; Ross Tomaino (Harvard Medical School) for mass spectrometry analysis; Y.L. Chong (Institute of Molecular and Cell Biology) for identification of zebrafish mutants; the A?STAR Microscopy Platform and the National University of Singapore EM unit for TEM analysis of zebrafish KV; staff at the Mary Lyon Centre for mouse husbandry, genotyping, and histology; Henrik Westerberg and Jenni Vibert (MRC Harwell Institute) for help with particle image velocimetry; Claire Jackson, Janice Coles, Liz Adams, and Peter Lackie (PCD Centre, University of Southampton) for assessing mouse tracheal ciliary beating and TEM images; Elspeth Bruford (HGNC) for gene nomenclature discussions; and Marie Bao for comments. P.A. and D.W.M. were supported by University of Sheffield-A?STAR Singapore and University of Manchester-A?STAR Singapore doctoral studentships, respectively. This work was supported by awards from the UK Medical Research Council (MC_U142670370 to D.P.N.), A?STAR Singapore (SC15-R0010 to S.R.), National Institutes of Health (NIGMS 1R01GM141109), the LouLou Foundation, the E. Matilda Ziegler Foundation for the Blind, the Smith Family Foundation, and the Pew Charitable Trusts (to A.B.). The National PCD Centre in Southampton is commissioned and funded by NHS England; PCD research is supported by the National Institute for Health Research (RfPB PB-PG-1215-20014 and 200470) and The AAIR Charity (Reg. No. 1129698). M.G. and J.J.B. purified samples, collected cryo-EM data, and picked particles with help from S.M. S.K.S. and Q.Z. M.G. processed the cryo-EM data. M.G. and J.R.A. built models and analyzed structures. H.F. and J.B.W. analyzed mouse situs and nodal and tracheal cilia motility. P.A. and D.W.M. created and characterized zebrafish mutants. T.Q. performed TEM of KV cilia. J.T. videoed and analyzed cilia motility and performed TEM of mouse tracheal cilia. J.S.L. C.D.B. D.P.N. S.R. and A.B. supervised the research. M.G. J.R.A. J.B.W. S.R. and A.B. wrote the manuscript with input from all authors. The authors declare no competing interests. Publisher Copyright: © 2021 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

Keywords: axonemal dyneins, ciliary motility, ciliopathies, cryo-EM, doublet microtubules, microtubule inner proteins

Local EPrints ID: 452211

URI: http://eprints.soton.ac.uk/id/eprint/452211

DOI: doi:10.1016/j.cell.2021.10.007

ISSN: 0092-8674

PURE UUID: 9a94c60a-3b66-4795-a406-2b6ab5c84db0

ORCID for James Thompson: orcid.org/0000-0002-9285-1317

ORCID for Jane Lucas: orcid.org/0000-0001-8701-9975

Date deposited: 30 Nov 2021 17:31

Last modified: 17 Mar 2024 03:32