title: TTC25 Deficiency Results in Defects of the Outer Dynein Arm Docking Machinery and Primary Ciliary Dyskinesia with Left-Right Body Asymmetry Randomization.
abstract: Multiprotein complexes referred to as outer dynein arms (ODAs) develop the main mechanical force to generate the ciliary and flagellar beat. ODA defects are the most common cause of primary ciliary dyskinesia (PCD), a congenital disorder of ciliary beating, characterized by recurrent infections of the upper and lower airways, as well as by progressive lung failure and randomization of left-right body asymmetry. Using a whole-exome sequencing approach, we identified recessive loss-of-function mutations within TTC25 in three individuals from two unrelated families affected by PCD. Mice generated by CRISPR/Cas9 technology and carrying a deletion of exons 2 and 3 in Ttc25 presented with laterality defects. Consistently, we observed immotile nodal cilia and missing leftward flow via particle image velocimetry. Furthermore, transmission electron microscopy (TEM) analysis in TTC25-deficient mice revealed an absence of ODAs. Consistent with our findings in mice, we were able to show loss of the ciliary ODAs in humans via TEM and immunofluorescence (IF) analyses. Additionally, IF analyses revealed an absence of the ODA docking complex (ODA-DC), along with its known components CCDC114, CCDC151, and ARMC4. Co-immunoprecipitation revealed interaction between the ODA-DC component CCDC114 and TTC25. Thus, here we report TTC25 as a new member of the ODA-DC machinery in humans and mice.
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Wallmeier
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Shiratori
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Gerard W
authors.initials: GW
authors.lastname: Dougherty
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Edelbusch
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Rim
authors.initials: R
authors.lastname: Hjeij
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Niki T
authors.initials: NT
authors.lastname: Loges
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Tabea
authors.initials: T
authors.lastname: Menchen
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Olbrich
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Pennekamp
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Raidt
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany.
authors.forename: Claudius
authors.initials: C
authors.lastname: Werner
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Katsura
authors.initials: K
authors.lastname: Minegishi
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Shinohara
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Asai
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Takaoka
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Chanjae
authors.initials: C
authors.lastname: Lee
authors.affiliation: Department of Pediatric Pulmonology, Hauner Children's Hospital and Ludwig Maximilian University, The German Center for Lung Research (DZL), 80337 Munich, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Griese
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
authors.forename: Yasin
authors.initials: Y
authors.lastname: Memari
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Durbin
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK.
authors.forename: Anja
authors.initials: A
authors.lastname: Kolb-Kokocinski
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK; Berlin Institute for Medical Systems Biology and Berlin Institute of Health Genomics Platforms, Laboratory of Functional Genomics, Nutrigenomics, and Systems Biology, Max Delbrück Center for Molecular Medicine, Robert Rössle Straße 10, 13125 Berlin, Germany; Otto Warburg Laboratory, Max Planck Institute for Molecular Genetics, Ihnestraße 63-73, 14195 Berlin, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Sauer
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: John B
authors.initials: JB
authors.lastname: Wallingford
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hamada
authors.affiliation: Department of Pediatrics, University Hospital Muenster, 48149 Muenster, Germany. Electronic address: heymut.omran@ukmuenster.de.
authors.forename: Heymut
authors.initials: H
authors.lastname: Omran
mesh_terms: D000818:Animals; D054468:Axoneme / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D002352:Carrier Proteins / Q000235:genetics*; D002923:Cilia / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D004398:Dyneins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D059472:Exome / Q000235:genetics; D005091:Exons / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005808:Genes, Recessive; D006801:Humans; D007619:Kartagener Syndrome / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D011485:Protein Binding; D014981:Xenopus; D029867:Xenopus Proteins / Q000172:deficiency / Q000235:genetics
chemical_list: D002352:Carrier Proteins; D004398:Dyneins; D029867:Xenopus Proteins; C572196:TTC25 protein, Xenopus; C000618383:ODAD4 protein, human; C000618391:TTC25 protein, mouse