Presentation is loading. Please wait.

Presentation is loading. Please wait.

CLPB Variants Associated with Autosomal-Recessive Mitochondrial Disorder with Cataract, Neutropenia, Epilepsy, and Methylglutaconic Aciduria  Carol Saunders,

Published byRobert King Modified over 5 years ago

Similar presentations

Presentation on theme: "CLPB Variants Associated with Autosomal-Recessive Mitochondrial Disorder with Cataract, Neutropenia, Epilepsy, and Methylglutaconic Aciduria  Carol Saunders,"— Presentation transcript:

1 CLPB Variants Associated with Autosomal-Recessive Mitochondrial Disorder with Cataract, Neutropenia, Epilepsy, and Methylglutaconic Aciduria  Carol Saunders, Laurie Smith, Flemming Wibrand, Kirstine Ravn, Peter Bross, Isabelle Thiffault, Mette Christensen, Andrea Atherton, Emily Farrow, Neil Miller, Stephen F. Kingsmore, Elsebet Ostergaard  The American Journal of Human Genetics  Volume 96, Issue 2, Pages (February 2015) DOI: /j.ajhg Copyright © 2015 The American Society of Human Genetics Terms and Conditions

2 Figure 1 Identification of CLPB Variants in Five Individuals
(A) DNA sequence analysis of CLPB shows the position of the homozygous c.803C>T variant in the subject compared to the control. (B) Analysis of cDNA encompassing the c.803C>T variant shows a band of normal size in individual 1. To assess the functional effect of the c.803C>T variant on splicing, RNA was extracted from fibroblasts and reverse transcribed to cDNA with the SuperScript II Reverse Transcriptase kit (Invitrogen), and PCR of a 381 bp cDNA fragment encompassing exons 4–8 was performed. To assess the carrier frequency for the c.803C>T variant, a TaqMan assay was developed (Applied Biosystems). The PCR conditions were: 10 μl Universal PCR Master Mix, 0.5 μl 40 × assay mix, and 20–100 ng DNA in a total volume of 20 μl. The PCR program was 95°C for 10 min, and 50 cycles at 92°C for 15 s and 60°C for 1 min. The samples were run on an ABI Prism 7000 and analyzed with ABI SDS software. (C) The alignment of the amino acid sequences of CLPB homologs in different vertebrate species shows the conservation of the mutated threonine at position 268. (D) A schematic representation of human CLPB (not to scale) shows the predicted domains and the position of the p.Thr268Met substitution. The following abbreviations are used: ANK, ankyrin repeat; CC, coiled-coil domain; AAA+, AAA+ ATPase; and D2-small, ClpB-D2-small. (E) DNA sequence analysis of CLPB DNA shows the position of the heterozygous c.961A>T variant in subject 5 compared to the control (left) and of the heterozygous c.1249C>T (right). The American Journal of Human Genetics  , DOI: ( /j.ajhg ) Copyright © 2015 The American Society of Human Genetics Terms and Conditions

3 Figure 2 Analysis of CLPB Protein Levels and In-Gel Activity of Complexes I and IV (A) In-gel enzyme activity in liver from individual 5 shows normal activity of complexes I and IV. (B) Analysis by SDS-PAGE of CLPB protein in different human tissues showing ubiquitous localization of CLPB. (C and D) Immunoblot analysis of CLPB in fibroblasts from individuals 3 and 4 and liver from subject 5 (D) shows absence of CLPB protein. In brief, mitochondrial protein was isolated from fibroblasts and liver as previously reported.23 The samples (25–30 cμg protein/lane) were run on a 12% SDS polyacrylamide gel and transferred to a PVDF membrane. The membrane was probed with a polyclonal antibody against CLPB (Atlas Antibodies) at a 1:1,000 dilution and developed with a 1:1,000 dilution of goat anti-rabbit antibody (Dako). An antibody against porin (Proteintech) was used as a loading control at a 1:1,000 dilution, and SUCLA2 was used as a reference at a 1:1,000 dilution. The secondary antibody was goat anti-mouse at a 1:1,000 dilution (Dako). The bands were visualized with the Supersignal West Pico and Femto substrates (Thermo Fisher Scientific) and MicroChemi imaging (DNR Bioimaging Systems). The American Journal of Human Genetics  , DOI: ( /j.ajhg ) Copyright © 2015 The American Society of Human Genetics Terms and Conditions

Download ppt "CLPB Variants Associated with Autosomal-Recessive Mitochondrial Disorder with Cataract, Neutropenia, Epilepsy, and Methylglutaconic Aciduria  Carol Saunders,"

Similar presentations

Previous Estimates of Mitochondrial DNA Mutation Level Variance Did Not Account for Sampling Error: Comparing the mtDNA Genetic Bottleneck in Mice and.

Previous Estimates of Mitochondrial DNA Mutation Level Variance Did Not Account for Sampling Error: Comparing the mtDNA Genetic Bottleneck in Mice and.
Tissue-Specific Reduction in Splicing Efficiency of IKBKAP Due to the Major Mutation Associated with Familial Dysautonomia  Math P. Cuajungco, Maire Leyne,

Tissue-Specific Reduction in Splicing Efficiency of IKBKAP Due to the Major Mutation Associated with Familial Dysautonomia  Math P. Cuajungco, Maire Leyne,
Loss-of-Function Mutations in RSPH1 Cause Primary Ciliary Dyskinesia with Central- Complex and Radial-Spoke Defects  Esther Kott, Marie Legendre, Bruno.

Loss-of-Function Mutations in RSPH1 Cause Primary Ciliary Dyskinesia with Central- Complex and Radial-Spoke Defects  Esther Kott, Marie Legendre, Bruno.
Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant

Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant
Novel Splice Variants of IL-33: Differential Expression in Normal and Transformed Cells  Hidetoshi Tsuda, Mayumi Komine, Masaru Karakawa, Takafumi Etoh,

Novel Splice Variants of IL-33: Differential Expression in Normal and Transformed Cells  Hidetoshi Tsuda, Mayumi Komine, Masaru Karakawa, Takafumi Etoh,
Microcytic anemia and hepatic iron overload in a child with compound heterozygous mutations in DMT1 (SCL11A2)‏ by Achille Iolascon, Maria d'Apolito, Veronica.

Microcytic anemia and hepatic iron overload in a child with compound heterozygous mutations in DMT1 (SCL11A2)‏ by Achille Iolascon, Maria d'Apolito, Veronica.
A Missense Mutation (R565W) in Cirhin (FLJ14728) in North American Indian Childhood Cirrhosis  Pierre Chagnon, Jacques Michaud, Grant Mitchell, Jocelyne.

A Missense Mutation (R565W) in Cirhin (FLJ14728) in North American Indian Childhood Cirrhosis  Pierre Chagnon, Jacques Michaud, Grant Mitchell, Jocelyne.
A Nonsense Mutation in COQ9 Causes Autosomal-Recessive Neonatal-Onset Primary Coenzyme Q10 Deficiency: A Potentially Treatable Form of Mitochondrial Disease 

A Nonsense Mutation in COQ9 Causes Autosomal-Recessive Neonatal-Onset Primary Coenzyme Q10 Deficiency: A Potentially Treatable Form of Mitochondrial Disease 
A Missense Mutation in PRPF6 Causes Impairment of pre-mRNA Splicing and Autosomal-Dominant Retinitis Pigmentosa  Goranka Tanackovic, Adriana Ransijn,

A Missense Mutation in PRPF6 Causes Impairment of pre-mRNA Splicing and Autosomal-Dominant Retinitis Pigmentosa  Goranka Tanackovic, Adriana Ransijn,
Eija Siintola, Meral Topcu, Nina Aula, Hannes Lohi, Berge A

Eija Siintola, Meral Topcu, Nina Aula, Hannes Lohi, Berge A
Differential HFE allele expression in hemochromatosis heterozygotes

Differential HFE allele expression in hemochromatosis heterozygotes
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,
The Molecular Basis of Malonyl-CoA Decarboxylase Deficiency

The Molecular Basis of Malonyl-CoA Decarboxylase Deficiency
Thiamine Pyrophosphokinase Deficiency in Encephalopathic Children with Defects in the Pyruvate Oxidation Pathway  Johannes A. Mayr, Peter Freisinger,

Thiamine Pyrophosphokinase Deficiency in Encephalopathic Children with Defects in the Pyruvate Oxidation Pathway  Johannes A. Mayr, Peter Freisinger,
Reccurrent F8 Intronic Deletion Found in Mild Hemophilia A Causes Alu Exonization  Yohann Jourdy, Alexandre Janin, Mathilde Fretigny, Anne Lienhart, Claude.

Reccurrent F8 Intronic Deletion Found in Mild Hemophilia A Causes Alu Exonization  Yohann Jourdy, Alexandre Janin, Mathilde Fretigny, Anne Lienhart, Claude.
A Novel Skeletal Dysplasia with Developmental Delay and Acanthosis Nigricans Is Caused by a Lys650Met Mutation in the Fibroblast Growth Factor Receptor.

A Novel Skeletal Dysplasia with Developmental Delay and Acanthosis Nigricans Is Caused by a Lys650Met Mutation in the Fibroblast Growth Factor Receptor.
Dihydrofolate Reductase Deficiency Due to a Homozygous DHFR Mutation Causes Megaloblastic Anemia and Cerebral Folate Deficiency Leading to Severe Neurologic.

Dihydrofolate Reductase Deficiency Due to a Homozygous DHFR Mutation Causes Megaloblastic Anemia and Cerebral Folate Deficiency Leading to Severe Neurologic.
Rat mesangial α-endosulfine

Rat mesangial α-endosulfine
Mutations in CYC1, Encoding Cytochrome c1 Subunit of Respiratory Chain Complex III, Cause Insulin-Responsive Hyperglycemia  Pauline Gaignard, Minal Menezes,

Mutations in CYC1, Encoding Cytochrome c1 Subunit of Respiratory Chain Complex III, Cause Insulin-Responsive Hyperglycemia  Pauline Gaignard, Minal Menezes,
Bulge- and Basal Layer-Specific Expression of Fibroblast Growth Factor-13 (FHF-2) in Mouse Skin  Mitsuko Kawano, Satoshi Suzuki, Masashi Suzuki, Junko.

Bulge- and Basal Layer-Specific Expression of Fibroblast Growth Factor-13 (FHF-2) in Mouse Skin  Mitsuko Kawano, Satoshi Suzuki, Masashi Suzuki, Junko.

Similar presentations

Ads by Google