C.-H. Huang, G.-J. Cheng, M.E. Reid, Y. Chen 

Slides:



Advertisements
Similar presentations
2470 bp 1891 bp WT bp 2314 bp A B Fig. S1. Verification with PCR amplification of the.
Advertisements

Lisa Edelmann, Raj K. Pandita, Bernice E. Morrow 
Alternative Splicing of a Novel Glycophorin Allele GPHe(GL) Generates Two Protein Isoforms in the Human Erythrocyte Membrane by Cheng-Han Huang, Olga O.
Mark M Metzstein, H.Robert Horvitz  Molecular Cell 
RHD gene deletion occurred in the Rhesus box
Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant
by Cheng-Han Huang, Ying Chen, Marion E. Reid, and Christine Seidl
Mutation Spectrum of the Survival of Motor Neuron 1 and Functional Analysis of Variants in Chinese Spinal Muscular Atrophy  Yu-jin Qu, Jin-li Bai, Yan-yan.
A Common Genetic Polymorphism (46 C to T Substitution) in the 5′-Untranslated Region of the Coagulation Factor XII Gene Is Associated With Low Translation.
Consequences of T‐DNA insertion on SWP expression in swp mutant.
Truncation of Glycoprotein (GP) IIIa (▵ ) Prevents Complex Formation With GPIIb: Novel Mutation in Exon 11 of GPIIIa Associated With Thrombasthenia.
Tracy Dixon-Salazar, Jennifer L. Silhavy, Sarah E. Marsh, Carrie M
RNAi Related Mechanisms Affect Both Transcriptional and Posttranscriptional Transgene Silencing in Drosophila  Manika Pal-Bhadra, Utpal Bhadra, James.
A Clinical Grade Sequencing-Based Assay for CEBPA Mutation Testing
Allan Award Lecture: On Jumping Fields and “Jumping Genes”
Brca1 Controls Homology-Directed DNA Repair
A Novel Alu-Like Element Rearranged in the Dystrophin Gene Causes a Splicing Mutation in a Family with X-Linked Dilated Cardiomyopathy  Alessandra Ferlini,
Rose-Anne Romano, Barbara Birkaya, Satrajit Sinha 
Andrea Gaedigk, Amanda K. Riffel, J. Steven Leeder 
Novel Mutations in the LAMC2 Gene in Non-Herlitz Junctional Epidermolysis Bullosa: Effects on Laminin-5 Assembly, Secretion, and Deposition  Daniele Castiglia,
A Novel Syndrome Combining Thyroid and Neurological Abnormalities Is Associated with Mutations in a Monocarboxylate Transporter Gene  Alexandra M. Dumitrescu,
Mutations in a Novel Gene with Transmembrane Domains Underlie Usher Syndrome Type 3  Tarja Joensuu, Riikka Hämäläinen, Bo Yuan, Cheryl Johnson, Saara.
A Gene Mutated in Nephronophthisis and Retinitis Pigmentosa Encodes a Novel Protein, Nephroretinin, Conserved in Evolution  Edgar Otto, Julia Hoefele,
Analysis of an exon 1 polymorphism of the B2 bradykinin receptor gene and its transcript in normal subjects and patients with C1 inhibitor deficiency 
Peter Ianakiev, Michael W
Molecular cloning of pms916 salt hypersensitive T-DNA mutant.
Structure of the GM2A Gene: Identification of an Exon 2 Nonsense Mutation and a Naturally Occurring Transcript with an In-Frame Deletion of Exon 2  Biao.
Molecular Analysis of the Cyclic AMP-Dependent Protein Kinase A (PKA) Regulatory Subunit 1A (PRKAR1A) Gene in Patients with Carney Complex and Primary.
Laminin-5 Mutational Analysis in an Italian Cohort of Patients with Junctional Epidermolysis Bullosa  Patrizia Posteraro, Naomi De Luca, Guerrino Meneguzzi,
Isolated 2-Methylbutyrylglycinuria Caused by Short/Branched-Chain Acyl-CoA Dehydrogenase Deficiency: Identification of a New Enzyme Defect, Resolution.
Haploinsufficiency for One COL3A1 Allele of Type III Procollagen Results in a Phenotype Similar to the Vascular Form of Ehlers-Danlos Syndrome, Ehlers-Danlos.
Imprinting at the SMPD1 Locus: Implications for Acid Sphingomyelinase–Deficient Niemann-Pick Disease  Calogera M. Simonaro, Jae-Ho Park, Efrat Eliyahu,
A Multi-Exonic BRCA1 Deletion Identified in Multiple Families through Single Nucleotide Polymorphism Haplotype Pair Analysis and Gene Amplification with.
A Homozygous Nonsense Mutation in Type XVII Collagen Gene (COL17A1) Uncovers an Alternatively Spliced mRNA Accounting for an Unusually Mild Form of Non-Herlitz.
J. H. D. Bassett, S. A. Forbes, A. A. J. Pannett, S. E. Lloyd, P. T
Kimberly C. Sippel, Rebecca E. Fraioli, Gary D. Smith, Mary E
A Presenilin-1 Truncating Mutation Is Present in Two Cases with Autopsy-Confirmed Early-Onset Alzheimer Disease  Carolyn Tysoe, Joanne Whittaker, John.
Genetic Background of Congenital Chloride Diarrhea in High-Incidence Populations: Finland, Poland, and Saudi Arabia and Kuwait  Pia Höglund, Mari Auranen,
Fabry Disease: Novel α-Galactosidase A 3′-Terminal Mutations Result in Multiple Transcripts Due to Aberrant 3′-End Formation  Makiko Yasuda, Junaid Shabbeer,
Michaela Floeth, Leena Bruckner-Tuderman 
Inherited Junctional Epidermolysis Bullosa in the German Pointer: Establishment of a Large Animal Model  Annabelle Capt, Flavia Spirito, Eric Guaguere,
The Origins of Hypertrophic Cardiomyopathy–Causing Mutations in Two South African Subpopulations: A Unique Profile of Both Independent and Founder Events 
Small Evolutionarily Conserved RNA, Resembling C/D Box Small Nucleolar RNA, Is Transcribed from PWCR1, a Novel Imprinted Gene in the Prader-Willi Deletion.
Deletion of the Cytoplasmatic Domain of BP180/Collagen XVII Causes a Phenotype with Predominant Features of Epidermolysis Bullosa Simplex  Marcel Huber,
A Novel Point Mutation Affecting the Tyrosine Kinase Domain of the TRKA Gene in a Family with Congenital Insensitivity to Pain with Anhidrosis  Shinichi.
A Mutation in the Variable Repeat Region of the Aggrecan Gene (AGC1) Causes a Form of Spondyloepiphyseal Dysplasia Associated with Severe, Premature.
Volume 10, Issue 1, Pages (January 2017)
Volume 54, Issue 1, Pages (July 1998)
Alternative Splicing in the α-Galactosidase A Gene: Increased Exon Inclusion Results in the Fabry Cardiac Phenotype  Satoshi Ishii, Shoichiro Nakao, Reiko.
Volume 58, Issue 2, Pages (August 2000)
Volume 25, Issue 2, Pages (February 2017)
Opitz G/BBB Syndrome in Xp22: Mutations in the MID1 Gene Cluster in the Carboxy- Terminal Domain  Karin Gaudenz, Erich Roessler, Nandita Quaderi, Brunella.
Identification of FOXP2 Truncation as a Novel Cause of Developmental Speech and Language Deficits  Kay D. MacDermot, Elena Bonora, Nuala Sykes, Anne-Marie.
Assessing the Functional Characteristics of Synonymous and Nonsynonymous Mutation Candidates by Use of Large DNA Constructs  A.M. Eeds, D. Mortlock, R.
Compound Heterozygosity for Novel Splice Site Mutations in the BPAG2/COL17A1 Gene Underlies Generalized Atrophic Benign Epidermolysis Bullosa  Leena Pulkkinen,
Mutation Analysis of the Entire PKD1 Gene: Genetic and Diagnostic Implications  Sandro Rossetti, Lana Strmecki, Vicki Gamble, Sarah Burton, Vicky Sneddon,
A Unique Point Mutation in the PMP22 Gene Is Associated with Charcot-Marie-Tooth Disease and Deafness  Margaret J. Kovach, Jing-Ping Lin, Simeon Boyadjiev,
Sang-Hyun Song, Chunhui Hou, Ann Dean  Molecular Cell 
Tracy Dixon-Salazar, Jennifer L. Silhavy, Sarah E. Marsh, Carrie M
Volume 93, Issue 1, Pages (April 1998)
MicroRNA Binding Sites in Arabidopsis Class III HD-ZIP mRNAs Are Required for Methylation of the Template Chromosome  Ning Bao, Khar-Wai Lye, M.Kathryn.
Contiguous Deletion of the X-Linked Adrenoleukodystrophy Gene (ABCD1) and DXS1357E: A Novel Neonatal Phenotype Similar to Peroxisomal Biogenesis Disorders 
Mutation of the Ca2+ Channel β Subunit Gene Cchb4 Is Associated with Ataxia and Seizures in the Lethargic (lh) Mouse  Daniel L Burgess, Julie M Jones,
Kit-Sing Au, Adelaide A. Hebert, E. Steve Roach, Hope Northrup 
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,
Volume 18, Issue 4, Pages (May 2005)
Identification of a New Splice Form of the EDA1 Gene Permits Detection of Nearly All X- Linked Hypohidrotic Ectodermal Dysplasia Mutations  Alex W. Monreal,
Allan Award Lecture: On Jumping Fields and “Jumping Genes”
Sequence Homology between 4qter and 10qter Loci Facilitates the Instability of Subtelomeric KpnI Repeat Units Implicated in Facioscapulohumeral Muscular.
K. Miura, M. Obama, K. Yun, H. Masuzaki, Y. Ikeda, S. Yoshimura, T
Presentation transcript:

Rhmod Syndrome: A Family Study of the Translation-Initiator Mutation in the Rh50 Glycoprotein Gene  C.-H. Huang, G.-J. Cheng, M.E. Reid, Y. Chen  The American Journal of Human Genetics  Volume 64, Issue 1, Pages 108-117 (January 1999) DOI: 10.1086/302215 Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 1 Southern blot analysis of RH and RHAG loci in the Rhmod family. Genomic DNAs were digested with restriction enzyme SphI, and the blots were hybridized with the Rh30 (left panel) and Rh50 (right panel) cDNA probes. The gene origin and exon content of various SphI bands assigned to the polymorphic region of RH (Huang et al. 1996) are denoted. Note the reduction of intensity in RHD-specific 8.9- and 1.2-kb bands and the presence of a ce-specific band in all members except the son. The blot probed with full-length Rh50 cDNA showed no significant difference between controls and Rhmod family members. Size markers (in kb) of HindIII-cleaved lambda-phage DNA are shown to the left of the gel panels. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 2 Inheritance and linkage of RH in the Rhmod family. Genotypes of four members were determined on the basis of the combined data from SphI RFLPs (fig. 1) and sequences of the corresponding transcripts. Rh-antigen phenotypes are shown below each family member: a plus sign (+) denotes that the individual is positive; a minus sign (−) denotes that the individual is negative. The Ce or ce gene (unblackened boxes) is placed upstream of the D gene (blackened boxes) (Carritt et al. 1997). “d” denotes a deletion of the D gene (horizontal line). The Rhmod proband (i.e., SM) and her husband are genotypically identical at RH, in that they both carry the DCe and dce haplotypes. The random assortment of the two haplotypes resulted in DCe/DCe and DCe/dce children. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 3 Amplification and analysis of the initiator mutation in the Rhmod family. A, DNA sequence profiles spanning the genomic region relevant to translation initiation. The genomic sequences encompassing exon 1 were amplified by primers 5P-1 and In-1a (see table 1) and then were directly sequenced. The nucleotide in the +3 position of initiation codon ATG is indicated by a downward-pointing arrow. The husband shows a normal sequence, whereas the Rhmod proband (i.e., SM) and her children are homozygous and heterozygous for the G→T transversion, respectively. B, SSCP analysis. The genomic products used for SSCP analysis were amplified by primers 5P-2 and Ex-1a. They were denatured by heating, were separated on an 8% polyacrylamide gel, and then were stained with silver nitrate. The normal and abnormal bands are denoted by arrows to the left and right of the gel panels, respectively. The results readily demonstrate the homozygosity and heterozygosity for the point mutation. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 4 Immunoblot analysis of Rh30 polypeptides and Rh50 glycoprotein in the RBC membrane. Equal amounts of RBC-membrane ghosts from controls and Rhmod family members were loaded and separated by PAGE under nonreducing conditions. A, Immunoblots probed with the monoclonal antibody 2D10 against Rh50. A broad diffuse band in the range of 36–50 kD is detectable in normal and Rhmod heterozygotes but is barely seen in the Rhmod homozygote (i.e., SM). B, Immunoblots probed with LOR-15C9, a monoclonal antibody specific for the RhD polypeptide only. A band of 30–32 kD is seen in all D+ individuals but not in either the D− control or the Rhmod homozygote. Thus, the RhD protein was severely deficient, although the RhD gene was structurally normal and actively transcribed in Rhmod. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 5 In vitro transcription–coupled translation assays of the wild-type and Rhmod Rh50 constructs. A, Diagrams of the two pairs of Rh50 constructs cloned into the pRc/CMV2 vector. The four constructs all start from −27A and are placed downstream of the T7 promoter. The sizes (in bp) of their coding sequences, from ATG (denoted by blackened diamonds [⧫]) to the last codon, is given, and the predicted length of their polypeptides is bracketed. “WT-L” and “WT-S” denote long and short wild-type constructs, respectively; “Rhmod-L” and “Rhmod-S” denote long and short mutant constructs, respectively. B, Analysis of in vitro–translation products. In each panel, equal amounts of linearized control and Rhmod plasmids were used. The apparent molecular size (in kD) of each band is given. Left, 15% SDS-PAGE of [35S]-Met–labeled products translated from the paired long constructs. Right, 20% SDS-PAGE of [35S]-Met–labeled products translated from the paired short constructs. Note the presence of a single band in WT-S and of two smaller bands in Rhmod-S. The dark band at the bottom is due to free [35S]-Met. C, Alignment of Kozak consensus (top) with the first four in-frame ATGs (boldface), at positions 1, 8, 16, and 44, and their flanking sequences in RHAG. The sequences surrounding ATG codons farther downstream, at positions 60 and 69, are not shown. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions

Figure 6 Model for the Rhmod phenotype caused by an incompletely penetrated initiator mutation in the RHAG gene. In the Rhmod proband (i.e., SM), the two copies of RHAG contain the same G→T transversion in the ATG initiation codon (circled “T”) and are denoted by horizontal unblackened boxes; her RH locus harbors structurally normal D, Ce, and ce genes that are actively expressed. As shown, the translation-initiator mutation alters neither gene transcription nor mRNA splicing; however, it leads to alternative translation initiation, by selective use of the downstream cryptic initiation-signal sequences, thus producing truncated Rh50 proteins with different N-terminal sequences, which are denoted by notches. Such products are aberrant in structure and may directly affect their contact with the Rh30 protein, via their N-terminal domains. They may also be defective in other posttranslational events, such as membrane insertion and cellular transport. These potential changes could largely decrease the formation of a stable Rh complex in the plasma membrane, although the RH locus itself harbors no mutation. The American Journal of Human Genetics 1999 64, 108-117DOI: (10.1086/302215) Copyright © 1999 The American Society of Human Genetics Terms and Conditions