RESUMO
Wilson disease (WD) is an autosomal recessive disorder of copper transport which map to chromosome 13q14.3. In pursuit of the WD gene, we developed yeast artificial chromosome and cosmid contigs, and microsatellite markers which span the WD gene region. Linkage disequilibrium and haplotype analysis of 115 WD families confined the disease locus to a single marker interval. A candidate cDNA clone was mapped to this interval which, as shown in the accompanying paper, is very likely the WD gene. Our haplotype and mutation analyses predict that approximately half of all WD mutations will be rare in the American and Russian populations.
Assuntos
Cromossomos Humanos Par 13 , Haplótipos/genética , Degeneração Hepatolenticular/genética , Sequência de Bases , Cosmídeos , Família , Feminino , Marcadores Genéticos , Biblioteca Genômica , Genótipo , Humanos , Desequilíbrio de Ligação , Masculino , Dados de Sequência Molecular , MutaçãoRESUMO
Wilson disease (WD) is an autosomal recessive disorder characterized by the toxic accumulation of copper in a number of organs, particularly the liver and brain. As shown in the accompanying paper, linkage disequilibrium & haplotype analysis confirmed the disease locus to a single marker interval at 13q14.3. Here we describe a partial cDNA clone (pWD) which maps to this region and shows a particular 76% amino acid homology to the Menkes disease gene, Mc1. The predicted functional properties of the pWD gene together with its strong homology to Mc1, genetic mapping data and identification of four independent disease-specific mutations, provide convincing evidence that pWD is the Wilson disease gene.
Assuntos
Adenosina Trifosfatases/genética , Proteínas de Transporte de Cátions , Cobre/metabolismo , Degeneração Hepatolenticular/genética , Síndrome dos Cabelos Torcidos/genética , Homologia de Sequência de Aminoácidos , Sequência de Aminoácidos , Sequência de Bases , ATPases Transportadoras de Cobre , Expressão Gênica , Haplótipos , Humanos , Desequilíbrio de Ligação , Dados de Sequência Molecular , MutaçãoRESUMO
Best macular dystrophy (BMD), also known as vitelliform macular dystrophy (VMD2; OMIM 153700), is an autosomal dominant form of macular degeneration characterized by an abnormal accumulation of lipofuscin within and beneath the retinal pigment epithelium cells. In pursuit of the disease gene, we limited the minimum genetic region by recombination breakpoint analysis and mapped to this region a novel retina-specific gene (VMD2). Genetic mapping data, identification of five independent disease-specific mutations and expression studies provide evidence that mutations within the candidate gene are a cause of BMD. The 3' UTR of the candidate gene contains a region of antisense complementarity to the 3' UTR of the ferritin heavy-chain gene (FTH1), indicating the possibility of antisense interaction between VMD2 and FTH1 transcripts.
Assuntos
Proteínas do Olho/genética , Degeneração Macular/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bestrofinas , Caenorhabditis elegans/genética , Canais de Cloreto , Mapeamento Cromossômico , Clonagem Molecular , Análise Mutacional de DNA , DNA Complementar , Feminino , Ferritinas/genética , Homologia de Genes , Humanos , Canais Iônicos , Masculino , Camundongos , Dados de Sequência Molecular , LinhagemRESUMO
Stargardt-like macular dystrophy (STGD3, MIM 600110) and autosomal dominant macular dystrophy (adMD) are inherited forms of macular degeneration characterized by decreased visual acuity, macular atrophy and extensive fundus flecks. Genetic mapping data suggest that mutations in a single gene may be responsible for both conditions, already known to bear clinical resemblance. Here we limit the minimum genetic region for STGD3 and adMD to a 0.6-cM interval by recombination breakpoint mapping and identify a single 5-bp deletion within the protein-coding region of a new retinal photoreceptor-specific gene, ELOVL4, in all affected members of STGD3 and adMD families. Bioinformatic analysis of ELOVL4 revealed that it has homology to a group of yeast proteins that function in the biosynthesis of very long chain fatty acids. Our results are therefore the first to implicate the biosynthesis of fatty acids in the pathogenesis of inherited macular degeneration.
Assuntos
Proteínas do Olho/genética , Genes Dominantes/genética , Degeneração Macular/genética , Proteínas de Membrana/genética , Deleção de Sequência/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Cromossomos Humanos Par 6/genética , Clonagem Molecular , Análise Mutacional de DNA , Éxons/genética , Proteínas do Olho/química , Proteínas do Olho/metabolismo , Feminino , Humanos , Hibridização In Situ , Íntrons/genética , Escore Lod , Macaca mulatta/genética , Degeneração Macular/patologia , Masculino , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Camundongos , Dados de Sequência Molecular , Linhagem , RNA Mensageiro/análise , RNA Mensageiro/genética , Retina/metabolismo , Retina/patologia , Alinhamento de SequênciaRESUMO
The expression of genes coding for alpha and alpha III isoforms of Na+,K+-ATPase alpha-subunit has been studied in human kidney, brain, thyroid and liver cells. The expression was shown to be subjected to a tissue-specific control and also depended on the developmental stage. The tissue-specific expression of genes coding for different isoforms of the catalytic subunit of Na+,K+-ATPase perhaps may be attributed to various functions of proteins belonging to this family.
Assuntos
Regulação da Expressão Gênica , Genes , ATPase Trocadora de Sódio-Potássio/genética , Adulto , Envelhecimento , Sequência de Aminoácidos , Sequência de Bases , Criança , Códon , Embrião de Mamíferos , Humanos , Isoenzimas/genética , Substâncias Macromoleculares , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , Especificidade de Órgãos , RNA Mensageiro/genéticaRESUMO
cDNAs complementary to pig kidney mRNAs coding for alpha- and beta-subunits of Na+,K+-ATPase were cloned and sequenced. Selective tryptic hydrolysis of the alpha-subunit within the membrane-bound enzyme and tryptic hydrolysis of the immobilized isolated beta-subunit were also performed. The mature alpha- and beta-subunits contain 1016 and 302 amino acid residues, respectively. Structural data on the peptides from extramembrane regions of the alpha-subunit and on glycopeptides of the beta-subunit underlie a model for the transmembrane arrangement of Na+,K+-ATPase polypeptide chains.
Assuntos
Medula Renal/enzimologia , ATPase Trocadora de Sódio-Potássio , Sequência de Aminoácidos , Animais , Sequência de Bases , Membrana Celular/enzimologia , Fenômenos Químicos , Físico-Química , DNA/genética , Bicamadas Lipídicas , Proteínas de Membrana , Hibridização de Ácido Nucleico , Fragmentos de Peptídeos , Poli A/genética , RNA/genética , RNA Mensageiro/genética , ATPase Trocadora de Sódio-Potássio/genética , SuínosRESUMO
Five different nucleotide sequences have been found in the human genome homologous to the gene of the alpha-subunit of Na+,K+-ATPase. A comparative analysis of the primary structure of these genes in the region 749-1328 (in coordinates of cDNA from the pig alpha-subunit) is presented.
Assuntos
Genes , Família Multigênica , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Recombinante , Humanos , Homologia de Sequência do Ácido NucleicoRESUMO
Intra-individual tissue-specific restriction fragment length polymorphism (RFLP) has been demonstrated in DNA isolated from different mammalian tissues using cDNAs of alpha- and beta-subunits of Na+,K+-ATPase as hybridization probes. We propose that the RFLPs could result from gene rearrangements in the gene loci for the alpha- and beta-subunits of Na+,K+-ATPase. The changes in restriction patterns have been shown to occur during embryonic development and tumor formation. In addition, the tissue specificity of the expression of different genes of the family of Na+,K+-ATPase genes and their low expression in tumor cells have been demonstrated.
Assuntos
Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , ATPase Trocadora de Sódio-Potássio/genética , Animais , DNA/genética , Feto/enzimologia , Regulação da Expressão Gênica , Humanos , Camundongos , Neoplasias/enzimologia , Hibridização de Ácido Nucleico , CoelhosRESUMO
The primary structure of the putative regulatory region of a gene of the Na+,K+-ATPase multigene family in the human genome has been determined. This region includes the first exon with all of the untranslatable sequence of mRNA and a dozen nucleotides, coding for the first four amino acids of the hypothetic precursor of the alpha+-subunit. The entire region comprises over 1400 bp. The possible role of specific nucleotide blocks within this region in comparison with other genes is discussed.
Assuntos
Genes Reguladores , Genes , Família Multigênica , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Cosmídeos , Humanos , Camundongos , Dados de Sequência Molecular , Coelhos , Mapeamento por Restrição , Homologia de Sequência do Ácido NucleicoRESUMO
The existence of a chromosome gene family containing at least one gene and one pseudogene was shown for the Na+,K+-ATPase beta-subunit. A partial structure of the beta 1-gene was determined, the coding part of which was completely homologous to cDNA of the Na+,K+-ATPase beta I-subunit from HeLa cells. The region encoding the putative protein transmembrane domain was shown to be bordered by two introns. The structure of a pseudogene (beta psi) was determined. This pseudogene is processed and contains multiple stop codons. Its homology to the beta I-subunit cDNA from HeLa cells is about 88%.
Assuntos
ATPase Trocadora de Sódio-Potássio/genética , Animais , Sequência de Bases , Células HeLa , Humanos , Proteínas de Membrana/genética , Dados de Sequência Molecular , Família Multigênica , Pseudogenes , Mapeamento por Restrição , Homologia de Sequência do Ácido Nucleico , SuínosRESUMO
The multigene family of human Na,K-ATPase is composed of 5 alpha-subunit genes, 3 of which were shown to encode the functionally active alpha 1, alpha 2 and alpha 3 isoforms of the catalytic subunits. This report describes the isolation, mapping and partial sequencing of the fourth gene (ATP1AL1) that was demonstrated here to be functionally active and expressed in human brain and kidney. Limited DNA sequencing of the ATP1AL1 exons allowed one to suggest that the gene probably encodes a new ion transport ATPase rather than an isoform of the Na,K-ATPase or the closely related H,K-ATPase.
Assuntos
Adenosina Trifosfatases/genética , Família Multigênica , ATPase Trocadora de Sódio-Potássio/genética , Transcrição Gênica , Adenosina Trifosfatases/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Transporte Biológico , Encéfalo/enzimologia , Eletroforese em Gel de Ágar , Éxons , Humanos , Íntrons , Rim/enzimologia , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , Reação em Cadeia da Polimerase , Homologia de Sequência do Ácido NucleicoRESUMO
PURPOSE: A 5-bp deletion in ELOVL4, a photoreceptor-specific gene, has been associated with autosomal dominant (ad) macular dystrophy phenotypes in five related families, in which phenotypes range from Stargardt-like macular dystrophy (STGD3; Mendelian Inheritance in Man 600110) to pattern dystrophy. This has been the only mutation identified in ELOVL4 to date, which is associated with macular dystrophy phenotypes. In the current study, the potential involvement was investigated of an ELOVL4 gene variation in adSTGD-like and other macular dystrophy phenotypes segregating in a large unrelated pedigree from Utah (K4175). METHODS: The entire open reading frame of the ELOVL4 gene was analyzed by direct sequencing in a proband from the K4175 family. The combination of denaturing high-performance liquid chromatography (DHPLC) analysis and direct sequencing of all available family members was used to further assess segregation of identified ELOVL4 variants in the pedigree. RESULTS: A complex mutation, two 1-bp deletions separated by four nucleotides, was detected in all affected members of the family. The mutation results in a frameshift and the truncation of the ELOVL4 protein, similar to the effect of the previously described 5-bp deletion. CONCLUSIONS: The discovery of a second mutation in the ELOVL4 gene segregating with macular dystrophy phenotypes confirms the role of this gene in a subset of dominant macular dystrophies with a wide range of clinical expressions and suggests a role for modifying genes and/or environmental factors in the disease process.
Assuntos
Proteínas do Olho/genética , Degeneração Macular/genética , Proteínas de Membrana/genética , Mutação/fisiologia , Adolescente , Adulto , Idoso , Criança , Feminino , Ligação Genética , Humanos , Degeneração Macular/patologia , Masculino , Pessoa de Meia-Idade , Linhagem , Penetrância , FenótipoRESUMO
Stargardt-like macular degeneration (STGD(3)) and autosomal dominant macular degeneration (adMD) share phenotypic characters with atrophic age-related macular degeneration (AMD). Mutations in a photoreceptor cell-specific factor involved in the elongation of very long chain fatty acids (ELOVL(4)) were shown to be associated with STGD(3), adMD, and pattern dystrophy. We screened 778 patients with AMD and 551 age-matched controls to define the role of sequence variants in the ELOVL(4) gene in age-related macular degeneration. We detected three sequence variants in the non-coding region and eight variants in the coding region. No statistically significant association was observed between sequence variants in the ELOVL(4) gene and susceptibility to AMD. However, for the detection of modest effects of multiple alleles in a complex disease, the analysis of larger cohorts of patients may be required.
Assuntos
Proteínas do Olho/genética , Degeneração Macular/genética , Proteínas de Membrana/genética , Mutação/genética , Polimorfismo de Nucleotídeo Único/genética , Fatores Etários , Idoso , Primers do DNA/química , Éxons , Variação Genética , Humanos , Íntrons , Degeneração Macular/fisiopatologia , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
mRNA coding for beta-subunit of Na+, K+-ATPase from pig kidney is about 24-25S as deduced from the hybridization pattern of poly (A)+-RNA with two synthetic oligonucleotides structurally corresponding to two peptides isolated from the tryptic hydrolyzate of beta-subunit. Cloning of cDNA allowed to determine the complete structure of the gene and to deduce the amino acid sequence of beta-subunit of Na+, K+-ATPase. The beta-subunit contains 302 amino acid residues and the protein is not processed at the N-nor at the C-terminus.
Assuntos
Genes , Rim/enzimologia , RNA Mensageiro/genética , DNA Polimerase Dirigida por RNA/genética , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA/genética , Hidrólise , Hibridização de Ácido Nucleico , Suínos , Moldes Genéticos , TripsinaRESUMO
Oligonucleotides deduced from the amino acid sequence of a hexapeptide Lys-Asp-Phe-Ala-Glu-Asn were synthesized and used as probes to screen a pig kidney cDNA library for a specific DNA sequence coding for the alpha-subunit of Na+, K+-ATPase. It was shown that the mixed oligoprobe, consisting of 64 heptadecamers, could be only suitable for mRNA blot analysis. To identify the clones with specific cDNA inserts, mixed oligoprobes were fractionated by HPLC technique. For the same purpose a new set of oligonucleotides, synthesized as four groups of 16 different heptadecamers each, was used.
Assuntos
DNA/genética , Genes , Oligonucleotídeos/genética , RNA Mensageiro/genética , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Medula Renal/enzimologia , Hibridização de Ácido Nucleico , Peptídeos/genética , ATPase Trocadora de Sódio-Potássio/análise , SuínosRESUMO
Messenger RNA, coding for the alpha-subunit of the Na+, K+-ATPase, was isolated from outer medulla of pig kidney. Within 25S-26S region the mRNA yields a band of specific hybridization with three oligonucleotide probes synthesized according to data on structures of three peptides isolated from the tryptic hydrolysate of the protein. Translation of the enriched poly(A+)-fraction of RNA in Xenopus laevis oocytes followed by the immunochemical identification of the products confirmed the presence of RNA coding for the desired protein. This RNA preparation was used for synthesis and cloning of double stranded cDNA.
Assuntos
Clonagem Molecular , Biossíntese de Proteínas , RNA Mensageiro/isolamento & purificação , DNA Polimerase Dirigida por RNA/genética , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , DNA/biossíntese , DNA/genética , Medula Renal/enzimologia , Hibridização de Ácido Nucleico , RNA Mensageiro/genética , ATPase Trocadora de Sódio-Potássio/análise , Suínos , Xenopus laevisRESUMO
The nucleotide sequence of the cDNA, containing coding region of the alpha-subunit of the pig kidney Na+, K+-ATPase, was determined. The region contains 3063 b.p. coding for 1021 amino acid residues. In the course of processing, five amino acid residues are cleaved to yield the mature Na+, K+-ATPase alpha-subunit containing 1016 amino acid residues.
Assuntos
Genes , Rim/enzimologia , ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , SuínosAssuntos
ATPase Trocadora de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA/análise , Enzimas de Restrição do DNA/metabolismo , Células HeLa/enzimologia , Humanos , Rim/enzimologia , Substâncias Macromoleculares , Hibridização de Ácido Nucleico , Ratos , Ovinos , SuínosAssuntos
Transportadores de Cassetes de Ligação de ATP , Adenosina Trifosfatases/biossíntese , Proteínas de Transporte/biossíntese , Proteínas de Transporte de Cátions , Proteínas de Escherichia coli , Proteínas de Transporte de Monossacarídeos , Adenosina Trifosfatases/química , Sequência de Aminoácidos , Sítios de Ligação , Proteínas de Transporte/química , Clonagem Molecular , Cobre/metabolismo , ATPases Transportadoras de Cobre , Escherichia coli , Degeneração Hepatolenticular/enzimologia , Humanos , Proteínas Ligantes de Maltose , Síndrome dos Cabelos Torcidos/enzimologia , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/químicaRESUMO
In this review we discuss four genetic disorders of copper metabolism. Wilson's disease and Indian childhood cirrhosis result from the toxic effects of copper accumulation in the liver. Menkes' disease and, most likely, occipital horn syndrome result from copper deficiency secondary to disturbances in copper transport. The recent cloning and sequencing of the genes defective in Wilson's disease and Menkes' disease provide the molecular basis for understanding the causes of the two major disorders of copper transport in humans. Mutations that result in Wilson's and Menkes' diseases were shown to disrupt the function of two related P-type copper transporting ATPases. Genetic analysis demonstrates that Wilson's disease and, probably, Menkes' disease are caused by a number of different mutations within a single gene (allelic heterogeneity), and that this occurrence likely explains the clinical heterogeneity of both diseases. The possibility that different mutations within the same gene account for the similar phenotypes of Wilson's disease and Indian childhood cirrhosis on the one hand and for Menkes' disease and occipital horn syndrome on the other are discussed.