title: Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease.
abstract: Ciliopathies are genetic disorders arising from dysfunction of microtubule-based cellular appendages called cilia. Different cilia types possess distinct stereotypic microtubule doublet arrangements with non-motile or 'primary' cilia having a 9+0 and motile cilia have a 9+2 array of microtubule doublets. Primary cilia are critical sensory and signaling centers needed for normal mammalian development. Defects in their structure/function result in a spectrum of clinical and developmental pathologies including abnormal neural tube and limb patterning. Altered patterning phenotypes in the limb and neural tube are due to perturbations in the hedgehog (Hh) signaling pathway. Motile cilia are important in fluid movement and defects in motility result in chronic respiratory infections, altered left-right asymmetry, and infertility. These features are the hallmarks of Primary Ciliary Dyskinesia (PCD, OMIM 244400). While mutations in several genes are associated with PCD in patients and animal models, the genetic lesion in many cases is unknown. We assessed the in vivo functions of Growth Arrest Specific 8 (GAS8). GAS8 shares strong sequence similarity with the Chlamydomonas Nexin-Dynein Regulatory Complex (NDRC) protein 4 (DRC4) where it is needed for proper flagella motility. In mammalian cells, the GAS8 protein localizes not only to the microtubule axoneme of motile cilia, but also to the base of non-motile cilia. Gas8 was recently implicated in the Hh signaling pathway as a regulator of Smoothened trafficking into the cilium. Here, we generate the first mouse with a Gas8 mutation and show that it causes severe PCD phenotypes; however, there were no overt Hh pathway phenotypes. In addition, we identified two human patients with missense variants in Gas8. Rescue experiments in Chlamydomonas revealed a subtle defect in swim velocity compared to controls. Further experiments using CRISPR/Cas9 homology driven repair (HDR) to generate one of these human missense variants in mice demonstrated that this allele is likely pathogenic.
authors.affiliation: Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Wesley R
authors.initials: WR
authors.lastname: Lewis
authors.affiliation: Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Erik B
authors.initials: EB
authors.lastname: Malarkey
authors.affiliation: Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Douglas
authors.initials: D
authors.lastname: Tritschler
authors.affiliation: Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Raqual
authors.initials: R
authors.lastname: Bower
authors.affiliation: Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Pasek
authors.affiliation: Division of Nephrology, Department of Medicine, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Porath
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.|Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Birket
authors.affiliation: INSERM, U-983, Necker Hospital, Paris, France.|Paris Descartes University, Sorbonne Paris Cité, Imagine Institute, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Saunier
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM) Unité Mixte de Recherche (UMR) 1163, Laboratory of Hereditary Kidney Diseases, Paris, France.|Assistance Publique-Hôpitaux de Paris, Department of Genetics, Necker Hospital, Paris, France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Antignac
authors.affiliation: Department of Medicine, UNC School of Medicine, Marisco Lung Institute, Chapel Hill, North Carolina, United States of America.
authors.forename: Michael R
authors.identifier: 0000-0002-4672-5969
authors.initials: MR
authors.lastname: Knowles
authors.affiliation: Department of Pediatrics, UNC School of Medicine, Marisco Lung Institute, Chapel Hill, North Carolina, United States of America.
authors.forename: Margaret W
authors.initials: MW
authors.lastname: Leigh
authors.affiliation: Department of Pathology and Laboratory Medicine, UNC School of Medicine, Marisco Lung Institute, Chapel Hill, North Carolina, United States of America.
authors.forename: Maimoona A
authors.initials: MA
authors.lastname: Zariwala
authors.affiliation: Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Anil K
authors.identifier: 0000-0002-3633-3304
authors.initials: AK
authors.lastname: Challa
authors.affiliation: Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Robert A
authors.identifier: 0000-0003-1331-0780
authors.initials: RA
authors.lastname: Kesterson
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.|Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Steven M
authors.identifier: 0000-0001-9045-0133
authors.initials: SM
authors.lastname: Rowe
authors.affiliation: Nephrology Division, Massachusetts General Hospital, Charlestown, Massachusetts, United States of America.
authors.forename: Iain A
authors.initials: IA
authors.lastname: Drummond
authors.affiliation: Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: John M
authors.initials: JM
authors.lastname: Parant
authors.affiliation: Division of Nephrology, Department of Medicine, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Hildebrandt
authors.affiliation: Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Mary E
authors.identifier: 0000-0001-8682-7655
authors.initials: ME
authors.lastname: Porter
authors.affiliation: Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Bradley K
authors.initials: BK
authors.lastname: Yoder
authors.affiliation: Department of Biology, Indiana University-Purdue University Indianapolis, Indianapolis, Indiana, United States of America.
authors.forename: Nicolas F
authors.identifier: 0000-0003-2863-1069
authors.initials: NF
authors.lastname: Berbari
mesh_terms: D000818:Animals; D019521:Body Patterning / Q000235:genetics*; D002465:Cell Movement / Q000235:genetics; D002696:Chlamydomonas / Q000235:genetics; D002923:Cilia / Q000235:genetics* / Q000473:pathology; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000235:genetics; D004195:Disease Models, Animal; D005121:Extremities / Q000254:growth & development / Q000473:pathology; D020022:Genetic Predisposition to Disease; D006801:Humans; D007619:Kartagener Syndrome / Q000235:genetics* / Q000473:pathology; D051379:Mice; D008870:Microtubules / Q000235:genetics; D009154:Mutation; D054259:Neural Tube / Q000254:growth & development / Q000473:pathology; D011506:Proteins / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics
chemical_list: D003598:Cytoskeletal Proteins; C452182:Gas8 protein, mouse; D011506:Proteins