Aideen M. McInerney-Leo, Miriam Schmidts, Claudio R. Cortés, Paul J

Slides:

Advertisements

Similar presentations

Splice-Site Mutations in the Axonemal Outer Dynein Arm Docking Complex Gene CCDC114 Cause Primary Ciliary Dyskinesia Alexandros Onoufriadis, Tamara Paff,

Advertisements

Mutations in CSPP1 Cause Primary Cilia Abnormalities and Joubert Syndrome with or without Jeune Asphyxiating Thoracic Dystrophy Karina Tuz, Ruxandra.

A Nonsense Mutation in the Human Homolog of Drosophila rogdi Causes Kohlschutter– Tonz Syndrome Adi Mory, Efrat Dagan, Barbara Illi, Philippe Duquesnoy,

Mutations in the Beta Propeller WDR72 Cause Autosomal-Recessive Hypomaturation Amelogenesis Imperfecta Walid El-Sayed, David A. Parry, Roger C. Shore,

Volume 11, Issue 8, Pages (May 2015)

A Missense Mutation in PRPF6 Causes Impairment of pre-mRNA Splicing and Autosomal-Dominant Retinitis Pigmentosa Goranka Tanackovic, Adriana Ransijn,

Mutations in Histone Acetylase Modifier BRPF1 Cause an Autosomal-Dominant Form of Intellectual Disability with Associated Ptosis Francesca Mattioli,

Eija Siintola, Meral Topcu, Nina Aula, Hannes Lohi, Berge A

Mutations in TPRN Cause a Progressive Form of Autosomal-Recessive Nonsyndromic Hearing Loss Yun Li, Esther Pohl, Redouane Boulouiz, Margit Schraders,

Undifferentiated Small Round Cell Sarcomas with Rare EWS Gene Fusions

A Truncating Mutation of CEP135 Causes Primary Microcephaly and Disturbed Centrosomal Function Muhammad Sajid Hussain, Shahid Mahmood Baig, Sascha Neumann,

Jiang Chen, Christine Laclef, Alejandra Moncayo, Elizabeth R

Mutations in AQP5, Encoding a Water-Channel Protein, Cause Autosomal-Dominant Diffuse Nonepidermolytic Palmoplantar Keratoderma Diana C. Blaydon, Lisbet K.

Depletion of primary cilia in articular chondrocytes results in reduced Gli3 repressor to activator ratio, increased Hedgehog signaling, and symptoms.

A Defect in the TUSC3 Gene Is Associated with Autosomal Recessive Mental Retardation Masoud Garshasbi, Valeh Hadavi, Haleh Habibi, Kimia Kahrizi, Roxana.

Kimberly A. Aldinger, Stephen J. Mosca, Martine Tétreault, Jennifer C

Volume 25, Issue 6, Pages (June 2017)

Volume 25, Issue 9, Pages (September 2017)

Mutations in the Gene Encoding IFT Dynein Complex Component WDR34 Cause Jeune Asphyxiating Thoracic Dystrophy Miriam Schmidts, Julia Vodopiutz, Sonia.

The Genetic Basis of Pachyonychia Congenita

Analysis of Rare APC Variants at the mRNA Level

Volume 19, Issue 19, Pages (October 2009)

Genetic variants in the ETV5 gene in fertile and infertile men with nonobstructive azoospermia associated with Sertoli cell–only syndrome Moira K. O'Bryan,

Identification and Rescue of Splice Defects Caused by Two Neighboring Deep-Intronic ABCA4 Mutations Underlying Stargardt Disease Silvia Albert, Alejandro.

Peter Ianakiev, Michael W

Volume 22, Issue 2, Pages (February 2014)

A Human Homologue of the Drosophila melanogaster diaphanous Gene Is Disrupted in a Patient with Premature Ovarian Failure: Evidence for Conserved Function.

Mohammad Rashel, Ninche Alston, Soosan Ghazizadeh

A Recurrent Missense Mutation in ZP3 Causes Empty Follicle Syndrome and Female Infertility Tailai Chen, Yuehong Bian, Xiaoman Liu, Shigang Zhao, Keliang.

Short Stature, Onychodysplasia, Facial Dysmorphism, and Hypotrichosis Syndrome Is Caused by a POC1A Mutation Ofer Sarig, Sagi Nahum, Debora Rapaport,

RBPJ Mutations Identified in Two Families Affected by Adams-Oliver Syndrome Susan J. Hassed, Graham B. Wiley, Shaofeng Wang, Ji-Yun Lee, Shibo Li, Weihong.

Gorab Is Required for Dermal Condensate Cells to Respond to Hedgehog Signals during Hair Follicle Morphogenesis Ying Liu, Elizabeth R. Snedecor, Yeon.

Clinical, Molecular, and Cellular Features of Non-Puerto Rican Hermansky–Pudlak Syndrome Patients of Hispanic Descent Carmelo Carmona-Rivera, Gretchen.

Mental Retardation and Abnormal Skeletal Development (Dyggve-Melchior-Clausen Dysplasia) Due to Mutations in a Novel, Evolutionarily Conserved Gene Daniel.

Mutations in CSPP1 Lead to Classical Joubert Syndrome

Mutation of Solute Carrier SLC16A12 Associates with a Syndrome Combining Juvenile Cataract with Microcornea and Renal Glucosuria Barbara Kloeckener-Gruissem,

Yuko Oda, Lizhi Hu, Vadim Bul, Hashem Elalieh, Janardan K

Volume 20, Issue 2, Pages (July 2017)

Autosomal-Dominant Woolly Hair Resulting from Disruption of Keratin 74 (KRT74), a Potential Determinant of Human Hair Texture Yutaka Shimomura, Muhammad.

Mutations in the DBP-Deficiency Protein HSD17B4 Cause Ovarian Dysgenesis, Hearing Loss, and Ataxia of Perrault Syndrome Sarah B. Pierce, Tom Walsh, Karen.

The Graded Response to Sonic Hedgehog Depends on Cilia Architecture

A Mutation in the Variable Repeat Region of the Aggrecan Gene (AGC1) Causes a Form of Spondyloepiphyseal Dysplasia Associated with Severe, Premature.

Biallelic Mutations in PATL2 Cause Female Infertility Characterized by Oocyte Maturation Arrest Biaobang Chen, Zhihua Zhang, Xiaoxi Sun, Yanping Kuang,

Biallelic SUN5 Mutations Cause Autosomal-Recessive Acephalic Spermatozoa Syndrome Fuxi Zhu, Fengsong Wang, Xiaoyu Yang, Jingjing Zhang, Huan Wu, Zhou.

Molecular Therapy - Nucleic Acids

Mutations in the Gene Encoding IFT Dynein Complex Component WDR34 Cause Jeune Asphyxiating Thoracic Dystrophy Miriam Schmidts, Julia Vodopiutz, Sonia.

Mutations in ARMC9, which Encodes a Basal Body Protein, Cause Joubert Syndrome in Humans and Ciliopathy Phenotypes in Zebrafish Julie C. Van De Weghe,

Assessing the Functional Characteristics of Synonymous and Nonsynonymous Mutation Candidates by Use of Large DNA Constructs A.M. Eeds, D. Mortlock, R.

A Defect in the TUSC3 Gene Is Associated with Autosomal Recessive Mental Retardation Masoud Garshasbi, Valeh Hadavi, Haleh Habibi, Kimia Kahrizi, Roxana.

Compound Heterozygosity for Novel Splice Site Mutations in the BPAG2/COL17A1 Gene Underlies Generalized Atrophic Benign Epidermolysis Bullosa Leena Pulkkinen,

Wook Lew Journal of Investigative Dermatology

Mental Retardation and Abnormal Skeletal Development (Dyggve-Melchior-Clausen Dysplasia) Due to Mutations in a Novel, Evolutionarily Conserved Gene Daniel.

A Lack of Birbeck Granules in Langerhans Cells Is Associated with a Naturally Occurring Point Mutation in the Human Langerin Gene Pauline Verdijk, Remco.

Mutations in CSPP1, Encoding a Core Centrosomal Protein, Cause a Range of Ciliopathy Phenotypes in Humans Ranad Shaheen, Hanan E. Shamseldin, Catrina M.

Mutations in POLR3A and POLR3B Encoding RNA Polymerase III Subunits Cause an Autosomal-Recessive Hypomyelinating Leukoencephalopathy Hirotomo Saitsu,

Mutations in the Gene Encoding Capillary Morphogenesis Protein 2 Cause Juvenile Hyaline Fibromatosis and Infantile Systemic Hyalinosis Sandra Hanks,

Deletions and Point Mutations of LRRC50 Cause Primary Ciliary Dyskinesia Due to Dynein Arm Defects Niki Tomas Loges, Heike Olbrich, Anita Becker-Heck,

Cerebral Cavernomas in a Family with Multiple Cutaneous and Uterine Leiomyomas Associated with a New Mutation in the Fumarate Hydratase Gene Elena Campione,

Identification of CC2D2A as a Meckel Syndrome Gene Adds an Important Piece to the Ciliopathy Puzzle Jonna Tallila, Eveliina Jakkula, Leena Peltonen,

Mutations in HPSE2 Cause Urofacial Syndrome

Mutations in NEXN, a Z-Disc Gene, Are Associated with Hypertrophic Cardiomyopathy Hu Wang, Zhaohui Li, Jizheng Wang, Kai Sun, Qiqiong Cui, Lei Song, Yubao.

Volume 3, Issue 3, Pages (March 1999)

Ciliary Abnormalities Due to Defects in the Retrograde Transport Protein DYNC2H1 in Short-Rib Polydactyly Syndrome Amy E. Merrill, Barry Merriman, Claire.

Human and Mouse Mutations in WDR35 Cause Short-Rib Polydactyly Syndromes Due to Abnormal Ciliogenesis Pleasantine Mill, Paul J. Lockhart, Elizabeth Fitzpatrick,

Epigenetic Allele Silencing Unveils Recessive RYR1 Mutations in Core Myopathies Haiyan Zhou, Martin Brockington, Heinz Jungbluth, David Monk, Philip Stanier,

Marshall Syndrome Associated with a Splicing Defect at the COL11A1 Locus Andrew J. Griffith, Leslie K. Sprunger, D. Alexa Sirko-Osadsa, George E. Tiller,

Exon Skipping in IVD RNA Processing in Isovaleric Acidemia Caused by Point Mutations in the Coding Region of the IVD Gene Jerry Vockley, Peter K. Rogan,

Identification of Novel pro-α2(IX) Collagen Gene Mutations in Two Families with Distinctive Oligo-Epiphyseal Forms of Multiple Epiphyseal Dysplasia Paul.

Mutations in Histone Acetylase Modifier BRPF1 Cause an Autosomal-Dominant Form of Intellectual Disability with Associated Ptosis Francesca Mattioli,

Identification and Functional Analysis of ZIC3 Mutations in Heterotaxy and Related Congenital Heart Defects Stephanie M. Ware, Jianlan Peng, Lirong Zhu,

Presentation transcript:

Short-Rib Polydactyly and Jeune Syndromes Are Caused by Mutations in WDR60 Aideen M. McInerney-Leo, Miriam Schmidts, Claudio R. Cortés, Paul J. Leo, Blanca Gener, Andrew D. Courtney, Brooke Gardiner, Jessica A. Harris, Yeping Lu, Mhairi Marshall, Peter J. Scambler, Philip L. Beales, Matthew A. Brown, Andreas Zankl, Hannah M. Mitchison, Emma L. Duncan, Carol Wicking The American Journal of Human Genetics Volume 93, Issue 3, Pages 515-523 (September 2013) DOI: 10.1016/j.ajhg.2013.06.022 Copyright © 2013 The American Society of Human Genetics Terms and Conditions

Figure 1 Clinical Features of Individuals from the SRP and Jeune Syndrome Families (A and B) X-rays of individuals (A) SKDP42.3 (II-1 in Figure 2A) and (B) SKDP42.4 (II-2 in Figure 2A), showing shortening of the ribs and long bones and bowing of the limbs. (C–F) Individual JATD76 (II-1 in Figure 2A) displays typical hallmarks of Jeune syndrome: mildly narrowed thorax (C), also shown in (E); polydactyly (D, arrow points to remnants of an extra digit subsequently removed from the right hand); handlebar clavicles (E, asterisks); and acetabular spurs (F, arrows). The American Journal of Human Genetics 2013 93, 515-523DOI: (10.1016/j.ajhg.2013.06.022) Copyright © 2013 The American Society of Human Genetics Terms and Conditions

Figure 2 Segregation of WDR60 Variants and the Predicted Effect on Protein (A) Pedigree structure and WDR60 genotypes for the Australian SRPS family (SKDP42: individual II-1 = SKDP42.3; II-2 = SKDP42.4) and the Spanish Jeune syndrome family (JATD76). Sequencing chromatograms of genomic DNA show examples of each of the WDR60 mutations. Asterisks mark mutated base; dotted line in JATD76 sequence marks the intron 13/exon 14 junction. (B) Agarose gel showing RT-PCR products amplified from the proband in family JATD76 (II-1) and control cDNA, using primers in exon 13 and 18, spanning the splice mutation predicted to result in skipping of exon 14 (see Figure S1 for schema). Abbreviations are as follows: M, marker; C, control. (C) Sequencing chromatogram of the JATD76 (II-1) cDNA sample showing the nondeleted (upper line) and exon-14-skipped (lower line) alleles. Arrow indicates exon 15 boundary with exon 14 (control allele) or 13 (mutated allele). Note the sequencing shown is in the reverse orientation. Primer sequences available on request. (D) Schematic of human WDR60 showing the main protein domains and the positions of the residues affected by the three mutations found in this study. The residue altered by the c.2246C>T (p.Thr749Met) missense mutation present in both families is conserved back to Drosophila. (Human ENST00000407559; Mouse ENSMUSG00000042050; Chicken ENSGALG00000006558; Zebrafish ENSDART00000109166; C. elegans c27F2.1; Drosophila FBgn0034095.) The American Journal of Human Genetics 2013 93, 515-523DOI: (10.1016/j.ajhg.2013.06.022) Copyright © 2013 The American Society of Human Genetics Terms and Conditions

Figure 3 WDR60 Localizes to the Base of the Cilium in Human Chondrocytes and Ciliogenesis Is Impaired in Fibroblast Cultures from Affected Individuals (A–E) C-28/I2 immortalized human chondrocytes were serum starved for 24 hr in DMEM+0.2% FBS prior to fixing in 4% paraformaldehyde (PFA). Cells were stained with antibodies to acetylated α-tubulin (green) and WDR60 (red), showing localization of WDR60 at the base of the cilium (marked with arrowheads in A and B; see Table S4 for details of antibodies used). Nuclei were stained with 4′,6-Diamidino-2-Phenylindole (DAPI; blue). Cells were imaged on an Olympus Deltavision IX71 inverted microscope with a 100× objective. High-power images of boxed regions in (C) shown in (D) and (E). Scale bars represent 10 μm in (A)–(C) and 5 μm in (D) and (E). (F–H) Graphs showing a decreased percentage of ciliated cells in fibroblasts prepared from affected individuals SKDP42.3 (F), SKDP42.4 (G), and JATD76 (II-1; H) versus control fibroblast lines from postnatal foreskin (F, G) or adult skin (H). Cells were cultured for a maximum of six passages and serum starved and fixed as above. Acetylated α-tubulin-stained cilia were counted manually and expressed as a percentage of the number of nuclei. Data shown are from a minimum of three independent experiments. For each experiment at least 100 cells were counted, the mean percentage of ciliated cells was calculated, and the mean values compared statistically by a Student’s t test (∗∗∗p < 0.0001) with Prism 5 (Graphpad) software. Error bars represent standard error of the mean (SEM). The American Journal of Human Genetics 2013 93, 515-523DOI: (10.1016/j.ajhg.2013.06.022) Copyright © 2013 The American Society of Human Genetics Terms and Conditions

Figure 4 Localization of IFT and Trafficking Proteins in WDR60 Mutant Fibroblasts Fibroblasts from individual SKDP42.3 and control fibroblast lines were serum starved as described (see legend to Figure 3), fixed, and stained for (A–C) IFT144 (red), (D–F; J–L) IFT88 (red), (G–I) GLI2 (red), and (J–L) ARL13b (green). In (A)–(I), cells were costained for acetylated α-tubulin and γ-tubulin (green). Nuclei were stained with DAPI (blue), indicating that in (A) two independent cells are shown. Localization in cells in the mutant cultures that don’t form a detectable primary cilium are shown in (B), (E), (H), and (K). Arrows point to accumulated proteins. In those mutant cells that form a cilium, all proteins analyzed show a similar localization to control cells as shown in (C), (F), (I), and (L). For IFT144 experiments (A–C) only, cells were treated with saponin (0.05% in PBS) for 1 min prior to fixation in 4% PFA for 10 min. In (D)–(I), cells were fixed in ice-cold MeOH for 5 min, in (J)–(L) in 4% PFA. Cells were imaged on an Olympus Deltavision IX71 inverted microscope with a 100× objective. Scale bars represent 5 μm. See Table S4 for details of antibodies used. The American Journal of Human Genetics 2013 93, 515-523DOI: (10.1016/j.ajhg.2013.06.022) Copyright © 2013 The American Society of Human Genetics Terms and Conditions