Presentation is loading. Please wait.

Presentation is loading. Please wait.

Exome Sequencing Identifies CCDC8 Mutations in 3-M Syndrome, Suggesting that CCDC8 Contributes in a Pathway with CUL7 and OBSL1 to Control Human Growth 

Published bySabine Baril Modified over 5 years ago

Similar presentations

Presentation on theme: "Exome Sequencing Identifies CCDC8 Mutations in 3-M Syndrome, Suggesting that CCDC8 Contributes in a Pathway with CUL7 and OBSL1 to Control Human Growth "— Presentation transcript:

1 Exome Sequencing Identifies CCDC8 Mutations in 3-M Syndrome, Suggesting that CCDC8 Contributes in a Pathway with CUL7 and OBSL1 to Control Human Growth  Dan Hanson, Philip G. Murray, James O'Sullivan, Jill Urquhart, Sarah Daly, Sanjeev S. Bhaskar, Leslie G. Biesecker, Mars Skae, Claire Smith, Trevor Cole, Jeremy Kirk, Kate Chandler, Helen Kingston, Dian Donnai, Peter E. Clayton, Graeme C.M. Black  The American Journal of Human Genetics  Volume 89, Issue 1, Pages (July 2011) DOI: /j.ajhg Copyright © 2011 The American Society of Human Genetics Terms and Conditions

2 Figure 1 Mutations in CCDC8 Cause 3-M Syndrome
(A) Individuals with 3-M syndrome who have CCDC8 mutations: i) 3M-2, ii) 3M-5. Their phenotype is indistinguishable from individuals with CUL7 or OBSL1 mutations. (B) Region of homozygosity on chromosome 19 common to three affected consanguineous individuals from different families. Analysis of SNP data from the Affymetrix Genome-Wide Human SNP Array 6.0 was performed by AutoSNPa. Black and yellow bars indicate homozygous and heterozygous SNP calls, respectively. Isolated heterozygous SNP calls within the larger homozygous region are likely to correspond to missed calls. The region of common homozygosity is flanked by recombinants at rs located at 45,373,227 bp and rs located at 53,270,701 bp, as shown by the red boxes. (C) Schematic of the third 3-M syndrome critical region and identity of CCDC8 mutations. Shown is a genetic map of chromosome 19 highlighting the critical region located between rs and rs and the relative location of CCDC8 within the region. CCDC8 is a single-exon gene encoding a protein of 538 amino acids, and bioinformatic analysis has indicated a coiled-coil domain located between residues 349–369 and 513–535 and an alanine-rich domain located between residues 299 and 471. Location and chromatograms of mutations in CCDC8 in individuals with 3-M syndrome are indicated by arrows. The American Journal of Human Genetics  , DOI: ( /j.ajhg ) Copyright © 2011 The American Society of Human Genetics Terms and Conditions

3 Figure 2 Molecular Characterization of CCDC8
(A) Immunoblot analysis of CCDC8 reveals loss of CCDC8 in an individual with 3-M syndrome and a CCDC8 mutation. HEK293 cells were transfected with a CCDC8 clone 24 hr before harvesting for both cytoplasmic and nuclear fractions. Human skin fibroblasts from controls and individuals with 3-M syndrome who have CUL7, OBSL1, or CCDC8 mutations were also harvested for cytoplasmic and nuclear fractions via standard protocols. Direct immunoblotting was performed for CCDC8 (Abnova) and loading was controlled by GAPDH (Santa-Cruz), demonstrating that individuals with 3-M syndrome who have either CUL7 or OBSL1 mutations have normal levels of CCDC8, whereas CCDC8 was undetectable in individual 3M-2. (B) RT-PCR of CCDC8 and GAPDH in a variety of adult tissues. The FirstChoice Human Total RNA Survey Panel (Ambion) was used for assessing CCDC8 expression in 20 different human tissues; CCDC8 mRNA was expressed widely but with low levels in spleen, skeletal muscle, small intestine, kidney, and liver. (C) Characterization of interactions among CUL7, OBSL1, and CCDC8. Immunoprecipitation analysis of the interactions between Myc-tagged CUL7 and V5-tagged OBSL1, V5-tagged OBSL1 and CCDC8, and CUL7 and CCDC8 in HEK293 cells. Plasmids expressing Myc-tagged CUL7, V5-tagged-OBSL1, and CCDC8 were used for transfection of HEK293 cells. The presence of recombinant proteins in each indicated transfection set was determined by direct immunoblot analysis of protein extracts. Interactions between Myc-tagged CUL7 and V5-tagged OBSL1, V5-tagged OBSL1 and CCDC8, and CUL7 and CCDC8 variants were analyzed by immunoprecipitation of extracts from each indicated transfection set with antibodies against V5 (AbDSerotec), c-Myc (Santa-Cruz), CUL7 (Santa-Cruz), or CCDC8 (Abnova), followed by capture with magnetic Dynabeads (Invitrogen). The precipitates were analyzed by immunoblotting with specific antibodies. The American Journal of Human Genetics  , DOI: ( /j.ajhg ) Copyright © 2011 The American Society of Human Genetics Terms and Conditions

Download ppt "Exome Sequencing Identifies CCDC8 Mutations in 3-M Syndrome, Suggesting that CCDC8 Contributes in a Pathway with CUL7 and OBSL1 to Control Human Growth "

Similar presentations

MLANA/MART1 and SILV/PMEL17/GP100 Are Transcriptionally Regulated by MITF in Melanocytes and Melanoma  Jinyan Du, Arlo J. Miller, Hans R. Widlund, Martin.

MLANA/MART1 and SILV/PMEL17/GP100 Are Transcriptionally Regulated by MITF in Melanocytes and Melanoma  Jinyan Du, Arlo J. Miller, Hans R. Widlund, Martin.
Keratins as Susceptibility Genes for End-Stage Liver Disease

Keratins as Susceptibility Genes for End-Stage Liver Disease
The Primordial Growth Disorder 3-M Syndrome Connects Ubiquitination to the Cytoskeletal Adaptor OBSL1  Dan Hanson, Philip G. Murray, Amit Sud, Samia A.

The Primordial Growth Disorder 3-M Syndrome Connects Ubiquitination to the Cytoskeletal Adaptor OBSL1  Dan Hanson, Philip G. Murray, Amit Sud, Samia A.
Identification of EpCAM as the Gene for Congenital Tufting Enteropathy

Identification of EpCAM as the Gene for Congenital Tufting Enteropathy
Whole-Exome Sequencing Identifies FAM20A Mutations as a Cause of Amelogenesis Imperfecta and Gingival Hyperplasia Syndrome  James O'Sullivan, Carolina C.

Whole-Exome Sequencing Identifies FAM20A Mutations as a Cause of Amelogenesis Imperfecta and Gingival Hyperplasia Syndrome  James O'Sullivan, Carolina C.
Mutations in AGBL1 Cause Dominant Late-Onset Fuchs Corneal Dystrophy and Alter Protein-Protein Interaction with TCF4  S. Amer Riazuddin, Shivakumar Vasanth,

Mutations in AGBL1 Cause Dominant Late-Onset Fuchs Corneal Dystrophy and Alter Protein-Protein Interaction with TCF4  S. Amer Riazuddin, Shivakumar Vasanth,
A Homozygous Missense Mutation in TGM5 Abolishes Epidermal Transglutaminase 5 Activity and Causes Acral Peeling Skin Syndrome  Andrew J. Cassidy, Maurice.

A Homozygous Missense Mutation in TGM5 Abolishes Epidermal Transglutaminase 5 Activity and Causes Acral Peeling Skin Syndrome  Andrew J. Cassidy, Maurice.
Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant

Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant
Tracy Dixon-Salazar, Jennifer L. Silhavy, Sarah E. Marsh, Carrie M

Tracy Dixon-Salazar, Jennifer L. Silhavy, Sarah E. Marsh, Carrie M
Mutations in Histone Acetylase Modifier BRPF1 Cause an Autosomal-Dominant Form of Intellectual Disability with Associated Ptosis  Francesca Mattioli,

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

Eija Siintola, Meral Topcu, Nina Aula, Hannes Lohi, Berge A
Mutations in NALCN Cause an Autosomal-Recessive Syndrome with Severe Hypotonia, Speech Impairment, and Cognitive Delay  Moeenaldeen D. Al-Sayed, Hamad.

Mutations in NALCN Cause an Autosomal-Recessive Syndrome with Severe Hypotonia, Speech Impairment, and Cognitive Delay  Moeenaldeen D. Al-Sayed, Hamad.
Mutations in TPRN Cause a Progressive Form of Autosomal-Recessive Nonsyndromic Hearing Loss  Yun Li, Esther Pohl, Redouane Boulouiz, Margit Schraders,

Mutations in TPRN Cause a Progressive Form of Autosomal-Recessive Nonsyndromic Hearing Loss  Yun Li, Esther Pohl, Redouane Boulouiz, Margit Schraders,
Cohen Syndrome Is Caused by Mutations in a Novel Gene, COH1, Encoding a Transmembrane Protein with a Presumed Role in Vesicle-Mediated Sorting and Intracellular.

Cohen Syndrome Is Caused by Mutations in a Novel Gene, COH1, Encoding a Transmembrane Protein with a Presumed Role in Vesicle-Mediated Sorting and Intracellular.
Structural and Functional Mutations of the Perlecan Gene Cause Schwartz-Jampel Syndrome, with Myotonic Myopathy and Chondrodysplasia  Eri Arikawa-Hirasawa,

Structural and Functional Mutations of the Perlecan Gene Cause Schwartz-Jampel Syndrome, with Myotonic Myopathy and Chondrodysplasia  Eri Arikawa-Hirasawa,
Expression and regulation of MD-2s.

Expression and regulation of MD-2s.
A Nonsense Mutation in DHTKD1 Causes Charcot-Marie-Tooth Disease Type 2 in a Large Chinese Pedigree  Wang-yang Xu, Ming-min Gu, Lian-hua Sun, Wen-ting.

A Nonsense Mutation in DHTKD1 Causes Charcot-Marie-Tooth Disease Type 2 in a Large Chinese Pedigree  Wang-yang Xu, Ming-min Gu, Lian-hua Sun, Wen-ting.
Exome Sequencing and Functional Analysis Identifies BANF1 Mutation as the Cause of a Hereditary Progeroid Syndrome  Xose S. Puente, Victor Quesada, Fernando G.

Exome Sequencing and Functional Analysis Identifies BANF1 Mutation as the Cause of a Hereditary Progeroid Syndrome  Xose S. Puente, Victor Quesada, Fernando G.
Mutations in PADI6 Cause Female Infertility Characterized by Early Embryonic Arrest  Yao Xu, Yingli Shi, Jing Fu, Min Yu, Ruizhi Feng, Qing Sang, Bo Liang,

Mutations in PADI6 Cause Female Infertility Characterized by Early Embryonic Arrest  Yao Xu, Yingli Shi, Jing Fu, Min Yu, Ruizhi Feng, Qing Sang, Bo Liang,
Mutations of the SYCP3 Gene in Women with Recurrent Pregnancy Loss

Mutations of the SYCP3 Gene in Women with Recurrent Pregnancy Loss

Similar presentations

Ads by Google