RESUMO
BACKGROUND: CHILD syndrome, a rare hereditary disorder of keratinization (MIM 308050, 300275), is the acronym proposed by Happle to name a rare entity, characterized by congenital hemidysplasia, icthyosiform nevus and limb defects, ranging from digital hypoplasia to icthyosiform nevus and ipsilateral limb defects, ranging from digital hypoplasia to complete amelia. PATIENTS AND METHODS: A 9-month-old female infant presented with skin and limb defects involving the right side of her body. Clinical and laboratory evaluation was performed, including DNA sequence analysis of the NSDHL gene. RESULTS: Our patient presented with some of the typical clinical characteristics of CHILD syndrome, i.e. two large erythematous plaques with sharp borders, covered with yellow, wax-like scaling, on the right axilla and on the right groin, dysplastic right hand and alopecia of the right occipital area. The diagnosis was confirmed by DNA screening analysis, that detected a missense mutation c.314C-->T;p-A105V, in the coding region of the NSDHL gene (exon4) of our patient. CONCLUSIONS: This is the first report of CHILD syndrome ever reported in Greece. We suggest that the diagnosis of the syndrome is important for patient information and genetic counselling.
Assuntos
3-Hidroxiesteroide Desidrogenases/genética , Eritema/genética , Deformidades Congênitas dos Membros/genética , Nevo/genética , Eritema/etnologia , Feminino , Grécia , Humanos , Lactente , Deformidades Congênitas dos Membros/etnologia , Mutação de Sentido Incorreto/genética , Nevo/etnologia , SíndromeRESUMO
The X-linked dominant CHILD syndrome (congenital hemidysplasia with ichthyosiform nevus and limb defects) is a rare developmental defect characterized by a strictly lateralized inflammatory nevus. In the majority of cases, the right side of the body is affected. Ipsilateral hypoplastic lesions may involve the brain, skeletal structures, lungs, heart or kidneys. We describe a case of CHILD syndrome involving the left side of the body. Absence of metacarpal, metatarsal and phalangeal bones of the left hand and foot resulted in oligodactyly, with only 3 fingers and 1 toe. An ipsilateral inflammatory epidermal nevus with hyperkeratosis, parakeratosis, acanthosis and perivascular lymphohistiocytic infiltrate was strictly confined to the left half of the patient's body. The phenotype was shown to be associated with a deletion of exons 6-8 of the X-linked NSDHL gene, confirming that CHILD syndrome is due to loss of function of an enzyme involved in cholesterol biosynthesis.
Assuntos
3-Hidroxiesteroide Desidrogenases/genética , Anormalidades Múltiplas/diagnóstico , Deleção Cromossômica , Cromossomos Humanos Par 6 , Eritrodermia Ictiosiforme Congênita/genética , Deformidades Congênitas dos Membros/diagnóstico , Anormalidades Múltiplas/genética , Sequência de Bases , Pré-Escolar , Cromossomos Humanos Par 8 , Análise Mutacional de DNA , Éxons/genética , Feminino , Seguimentos , Humanos , Hidroxiesteroide Desidrogenases/genética , Eritrodermia Ictiosiforme Congênita/diagnóstico , Deformidades Congênitas dos Membros/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , SíndromeAssuntos
3-Hidroxiesteroide Desidrogenases/genética , Ictiose/genética , Deformidades Congênitas dos Membros/genética , Mutação , Nevo/genética , Adolescente , Sequência de Aminoácidos , Análise Mutacional de DNA , Feminino , Humanos , Ictiose/diagnóstico , Deformidades Congênitas dos Membros/diagnóstico , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Nevo/diagnóstico , Mutação Puntual , Polimorfismo de Nucleotídeo Único , Alinhamento de Sequência , Deleção de SequênciaAssuntos
Proteínas de Ligação a DNA/genética , Hamartoma/genética , Doenças Hipotalâmicas/genética , Proteínas do Tecido Nervoso , Fatores de Transcrição/genética , Sequência de Bases , Códon/genética , Códon sem Sentido , Humanos , Fatores de Transcrição Kruppel-Like , Síndrome , Proteína Gli3 com Dedos de ZincoRESUMO
Greig cephalopolysyndactyly (GCPS) (OMIM 175700) is an autosomal dominant disorder characterized by a distinct combination of craniofacial, hand and foot malformations. In this report, clinical and radiological findings of 12 patients with GCPS derived from 4 independent families and 3 sporadic cases with documented GLI3 mutations are presented with particular emphasis on inter- and intrafamilial variability. In a particularly instructive family in which 9 members of 4 generations could be studied clinically and molecularly, a missense mutation (R625W) is transmitted and shows a partially penetrant pattern. In a branch of the family, the GCPS phenotype skips a generation via a normal female carrier without clinical signs providing evidence that GCPS does not always manifest full penetrance as generally supposed.
Assuntos
Proteínas de Ligação a DNA/genética , Mutação , Proteínas do Tecido Nervoso , Fatores de Transcrição/genética , Alelos , Anormalidades Craniofaciais/diagnóstico por imagem , Anormalidades Craniofaciais/genética , Análise Mutacional de DNA , Fácies , Saúde da Família , Feminino , Genes Dominantes , Marcadores Genéticos , Heterozigoto , Humanos , Hibridização in Situ Fluorescente , Fatores de Transcrição Kruppel-Like , Deformidades Congênitas dos Membros/diagnóstico por imagem , Deformidades Congênitas dos Membros/genética , Masculino , Mutação de Sentido Incorreto , Linhagem , Penetrância , Fenótipo , Polidactilia/diagnóstico por imagem , Polidactilia/genética , Polimorfismo Conformacional de Fita Simples , Radiografia , Sindactilia/diagnóstico por imagem , Sindactilia/genética , Síndrome , Proteína Gli3 com Dedos de ZincoRESUMO
A technique for the Selective Differential Display of RNAs containing Interspersed Repeats (SDDIR) has been elaborated. SDDIR involves two main steps: (1) selective amplification by RT-PCR of a subset of the total cellular RNA containing a certain type of repetitive element, and (2) side-by-side display of the amplicons derived from the tissues under comparison by means of gel electrophoresis in parallel lanes. The technique was used to compare the expression of transcripts containing LTR (Long Terminal Repeat) sequences derived from human endogenous retrovirus K (HERV-K) in testicular germ cell tumors and in corresponding normal tissue. SDDIR enabled us to obtain an overview of LTRs represented in the total transcribed fraction and to reveal differences in transcription patterns of the LTRs in normal and tumor tissues. An unexpectedly large number of LTRs was found to be transcribed, and the levels of many of the transcripts differed between normal and tumor tissues.
Assuntos
Retrovirus Endógenos/genética , Germinoma/genética , Germinoma/virologia , RNA Neoplásico/genética , RNA Viral/genética , Neoplasias Testiculares/genética , Neoplasias Testiculares/virologia , Sequência de Bases , Expressão Gênica , Perfilação da Expressão Gênica , Humanos , Sequências Repetitivas Dispersas , Íntrons , Masculino , Seminoma/genética , Transcrição GênicaRESUMO
TFIIIB, TFIIIC2, and PTF/SNAPC are heteromultimeric general transcription factors (GTFs) needed for expression of genes encoding small cytoplasmic (scRNAs) and small nuclear RNAs (snRNAs). Their activity is stimulated by viral oncogenes, such as SV40 large T antigen and Adenovirus E1A, and is repressed by specific transcription factors (STFs) acting as anti-oncogenes, such as p53 and pRb. GTFs role as final targets of critical signal transduction pathways, that control cell proliferation and differentiation, and their involvement in gene expression regulation suggest that the genes encoding them are potential proto-oncogenes or anti-oncogenes or may be otherwise involved in the pathogenesis of inherited genetic diseases. To test our hypothesis through the positional candidate gene approach, we have determined the physical localization in the human genome of the 11 genes, encoding the subunits of these GTFs, and of three genes for proteins associated with TFIIIB (GTF3BAPs). Our data, obtained by chromosomal in situ hybridization, radiation hybrids and somatic cell hybrids analysis, demonstrate that these genes are present in the human genome as single copy sequences and that some cluster to the same cytogenetic band, alone or in combination with class II GTFs. Intriguingly, some of them are localized within chromosomal regions where recurrent, cytogenetically detectable mutations are seen in specific neoplasias, such as neuroblastoma, uterine leyomioma, mucoepidermoid carcinoma of the salivary glands and hemangiopericytoma, or where mutations causing inherited genetic diseases map, such as Peutz-Jeghers syndrome. Their molecular function and genomic position make these GTF genes interesting candidates for causal involvement in oncogenesis or in the pathogenesis of inherited genetic diseases.
Assuntos
Predisposição Genética para Doença , Genoma Humano , Neoplasias/etiologia , Fatores de Transcrição TFIII/genética , Fatores de Transcrição/genética , Mapeamento Cromossômico , Humanos , Neoplasias/genética , Fator de Transcrição TFIIIBRESUMO
An unusual combination of syndactylies, macrocephaly, and severe skeletal dysplasia was observed in a newborn infant. A history of digital anomalies in the father and grandfather lead to the diagnosis of dominantly inherited Greig cephalopolysyndactyly syndrome (GCPS, MIM #175700). Having explained the digital findings and macrocephaly, the skeletal changes were thought to fit best congenital spondyloepiphyseal dysplasia (SEDC MIM #183900), a type II collagen disorder. Molecular analysis confirmed the presence of two dominant mutations in the propositus: a GLI3 mutation (E543X), which was present also in the father and grandfather, and a de novo COL2A1 mutation leading to a G973R substitution. Thus, this boy combined the syndactyly-macrocephaly phenotype of Greig cephalosyndactyly syndrome with a severe form of spondyloepiphyseal dysplasia caused by the structural defect in type II collagen. The diagnostic difficulties posed by the combination of two genetic disorders and the contribution of molecular diagnostics are well illustrated by this case.
Assuntos
Anormalidades Múltiplas/genética , Colágeno/genética , Proteínas de Ligação a DNA/genética , Genes Dominantes , Proteínas do Tecido Nervoso , Mutação Puntual , Proteínas Repressoras , Fatores de Transcrição/genética , Proteínas de Xenopus , Feminino , Humanos , Lactente , Fatores de Transcrição Kruppel-Like , Escore Lod , Masculino , Osteocondrodisplasias/genética , Linhagem , Crânio/anormalidades , Proteína Gli3 com Dedos de ZincoRESUMO
We report for the first time that CHILD syndrome (MIM 308050), an X-linked dominant, male-lethal trait characterized by an inflammatory nevus with striking lateralization and strict midline demarcation, as well as ipsilateral hypoplasia of the body is caused by mutations in the gene NSDHL located at Xq28 (NAD(P)H steroid dehydrogenase-like protein) encoding a 3beta-hydroxysteroid dehydrogenase functioning in the cholesterol biosynthetic pathway. SSCA and genomic sequence analysis of NSDHL identified in 6 patients with CHILD syndrome, including one boy as well as a mother and her daughter, mutations potentially impairing protein function. This phenotype is distinct from, but shares various clinical and biochemical findings with chondrodysplasia punctata (CDPX2, MIM 302960). CDPX2 is due to mutations affecting a delta8-delta7 sterol isomerase (EBP, emopamil binding protein, at Xp11.22-p11.23) that functions downstream of NSDHL in a later step of cholesterol biosynthesis. EBP was unaffected in the patients analyzed by us demonstrating that CHILD syndrome and CDPX2 are not caused by allelic mutations. Two mouse X-linked dominant male-lethal traits, bare patches (Bpa) and striated (Str) had previously been associated with mutations in Nsdhl. They provide animal models for the study of CHILD syndrome, a further human condition due to mutations in a gene of the cholesterol synthesis pathway.
Assuntos
3-Hidroxiesteroide Desidrogenases/genética , Anormalidades Múltiplas/genética , Mutação , Anormalidades Múltiplas/enzimologia , Sequência de Aminoácidos , Sequência de Bases , DNA , Feminino , Humanos , Ictiose Ligada ao Cromossomo X/enzimologia , Ictiose Ligada ao Cromossomo X/genética , Deformidades Congênitas dos Membros/enzimologia , Deformidades Congênitas dos Membros/genética , Masculino , Dados de Sequência Molecular , SíndromeRESUMO
Greig cephalopolysyndactyly syndrome, characterized by craniofacial and limb anomalies (GCPS; MIM 175700), previously has been demonstrated to be associated with translocations as well as point mutations affecting one allele of the zinc finger gene GLI3. In addition to GCPS, Pallister-Hall syndrome (PHS; MIM 146510) and post-axial polydactyly type A (PAP-A; MIM 174200), two other disorders of human development, are caused by GLI3 mutations. In order to gain more insight into the mutational spectrum associated with a single phenotype, we report here the extension of the GLI3 mutation analysis to 24 new GCPS cases. We report the identification of 15 novel mutations present in one of the patient's GLI3 alleles. The mutations map throughout the coding gene regions. The majority are truncating mutations (nine of 15) that engender prematurely terminated protein products mostly but not exclusively N-terminally to or within the central region encoding the DNA-binding domain. Two missense and two splicing mutations mapping within the zinc finger motifs presumably also interfere with DNA binding. The five mutations identified within the protein regions C-terminal to the zinc fingers putatively affect additional functional properties of GLI3. In cell transfection experiments using fusions of the DNA-binding domain of yeast GAL4 to different segments of GLI3, transactivating capacity was assigned to two adjacent independent domains (TA(1)and TA(2)) in the C-terminal third of GLI3. Since these are the only functional domains affected by three C-terminally truncating mutations, we postulate that GCPS may be due either to haploinsufficiency resulting from the complete loss of one gene copy or to functional haploinsufficiency related to compromised properties of this transcription factor such as DNA binding and transactivation.
Assuntos
Anormalidades Craniofaciais/genética , Proteínas de Ligação a DNA/genética , Deformidades Congênitas dos Membros/genética , Mutação , Proteínas do Tecido Nervoso , Proteínas Repressoras , Fatores de Transcrição/genética , Proteínas de Xenopus , Animais , Análise Mutacional de DNA , Drosophila , Humanos , Fatores de Transcrição Kruppel-Like , Proteínas Recombinantes de Fusão , Deleção de Sequência , Síndrome , Ativação Transcricional , Transfecção , Células Tumorais Cultivadas , Proteína Gli3 com Dedos de Zinco , Dedos de Zinco/genéticaRESUMO
Functional characterization of a gene often requires the discovery of the full spectrum of its associated phenotypes. Mutations in the human GLI3 gene have been identified in Greig cepalopolysyndactyly, Pallister-Hall syndrome (PHS), and postaxial polydactyly type-A (PAP-A). We studied the involvement of GLI3 in additional phenotypes of digital abnormalities in one family (UR003) with preaxial polydactyly type-IV (PPD-IV), three families (UR014, UR015, and UR016) with dominant PAP-A/B (with PPD-A and -B in the same family), and one family with PHS. Linkage analysis showed no recombination with GLI3-linked polymorphisms. Family UR003 had a 1-nt frameshift insertion, resulting in a truncated protein of 1,245 amino acids. A frameshift mutation due to a 1-nt deletion was found in family UR014, resulting in a truncated protein of 1,280 amino acids. Family UR015 had a nonsense mutation, R643X, and family UR016 had a missense mutation, G727R, in a highly conserved amino acid of domain 3. The patient with PHS had a nonsense mutation, E1147X. These results add two phenotypes to the phenotypic spectrum caused by GLI3 mutations: the combined PAP-A/B and PPD-IV. These mutations do not support the suggested association between the mutations in GLI3 and the resulting phenotypes. We propose that all phenotypes associated with GLI3 mutations be called "GLI3 morphopathies," since the phenotypic borders of the resulting syndromes are not well defined and there is no apparent genotype-phenotype correlation.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Genes Dominantes/genética , Mutação , Proteínas do Tecido Nervoso , Polidactilia/genética , Proteínas Repressoras , Fatores de Transcrição/metabolismo , Proteínas de Xenopus , Sequência de Aminoácidos , Sequência de Bases , Cromossomos Humanos Par 7/genética , Códon/genética , Análise Mutacional de DNA , Proteínas de Ligação a DNA/genética , Éxons/genética , Saúde da Família , Feminino , Ligação Genética/genética , Genótipo , Humanos , Índia , Fatores de Transcrição Kruppel-Like , Masculino , Dados de Sequência Molecular , Linhagem , Fenótipo , Polidactilia/fisiopatologia , Polimorfismo Genético/genética , Síndrome , Fatores de Transcrição/genética , Proteína Gli3 com Dedos de ZincoRESUMO
By in situ chromosomal hybridization, and by somatic cell and radiation hybrid analysis, we have determined the genomic position of the human genes encoding four TAFII subunits of TFIID (TAFII150, TAFII105, TAFII68, TAFII18), the three subunits of TFIIA (TFIIA35 and TFIIA19, both encoded by the same gene, and TFIIA12), CDK8, and SURB7. All of these proteins are bona fide components of human class II holoenzymes as well as targets of signal transduction pathways that regulate genome expression. The genes encoding them are present in the human genome in a single copy and are localized at 8q23, 18q11.2, 17q11.1-11.2, 1p21, 14q31, 15q21-23, 13q12, and 12p12, respectively. We have mapped all of them to chromosomal regions where hereditary genetic diseases have been localized or which are involved in malignancies, which makes them potential candidates for a causal involvement in these phenotypes.
Assuntos
Quinases Ciclina-Dependentes , Proteínas de Ligação a DNA/genética , Proteínas Serina-Treonina Quinases/genética , Fatores de Transcrição TFII/genética , Fatores de Transcrição/genética , Transcrição Gênica/genética , Anormalidades Múltiplas/genética , Cromossomos Humanos , Quinase 8 Dependente de Ciclina , Dosagem de Genes , Genoma Humano , Humanos , Hibridização In Situ , Complexo Mediador , Mapeamento de Híbridos Radioativos , Fator de Transcrição TFIIA , Fator de Transcrição TFIIDRESUMO
We have isolated and characterized a human genomic clone containing the complete coding region of connexin31 (Cx31). Similar to rodent Cx31, the coding region of human Cx31 is completely contained within the second exon and consists of 810 nucleotides. The deduced human Cx31 polypeptide consists of 270 amino acids with a predicted molecular mass of 30.818 kDa. Its sequence is most similar to mouse Cx31 (82.6% identical amino acids) and rat (83.0% identical amino acids), but shows considerably fewer potential sites of phosphorylation. After Northern blot hybridization, two Cx31 transcripts of 2.2 and 1.8 kb were detected in total RNA of the human keratinocyte cell line HaCaT and two transcripts of 2.2 and 1.9 kb in total RNA of E6/E7 transfected human keratinocytes (HEK cells). Using affinity-purified rabbit antibodies to mouse Cx31, immunofluorescence analysis demonstrated relatively weak expression of human Cx31 in HaCaT and HEK cells. The Cx31 gene exists as a single copy gene in the human genome and was mapped to the chromosomal region 1p34-p36 by analyzing human-mouse somatic cell hybrids.
Assuntos
Conexinas/biossíntese , Conexinas/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Conexinas/genética , Feminino , Biblioteca Genômica , Células HeLa , Humanos , Camundongos , Dados de Sequência Molecular , Placenta , Gravidez , Coelhos , Ratos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , TransfecçãoRESUMO
TFIID, a multisubunit protein comprised of TBP (TATA box-binding protein) and TAF(II)s (TBP-associated factors), has a central role in transcription initiation at class II promoters. TAF(II)s role as mediators of regulatory transcription factors, such as pRb and p53, and their involvement in signal transduction pathways suggest that some may participate in the control of cell proliferation and differentiation: therefore, they could be considered potential protooncogenes or antioncogenes. With the aim of starting to analyse these potential roles, we have determined the genomic position of nine human TAF(II) genes (TAF[II]250, TAF[II]135, TAF[II]100, TAF[II]80, TAF[II]55, TAF[II]43, TAF[II]31, TAF[II]28, TAF[II]20/15) and of two previously unknown sequences related to TAF(II)250 and TAF(II)31, respectively. Except for those encoding TAF(II)250 and TAF(II)31, these genes are present in a single copy and, with the exclusion of those for TAF(II)43 and TAF(II)28 (both at 6p21), are localized in different segments of the genome. Indeed, six of them map to a chromosomal region commonly altered in specific neoplasias, which defines them as candidates for involvement in oncogenesis. Our experiments also demonstrate that TAF(II) transcripts are synthesized ubiquitously, mostly at low levels similar to those of TBP. Interestingly, the amount of the major mRNA species detected by TAF(II)20/15 cDNA is higher, which suggests that the polypeptide it encodes may also perform functions independently of TFIID. TAF(II) isoforms, indicated by additional bands on Northern blots, may play a role in modulation of TFIID function. These data will be useful for analysing variations of TAF(II) mRNA phenotype during cell proliferation, differentiation and development, both normal and pathological.
Assuntos
Genoma Humano , Fatores de Transcrição TFII/genética , Transcrição Gênica , Animais , Northern Blotting , Mapeamento Cromossômico , Humanos , Hibridização in Situ Fluorescente , Camundongos , RNA Mensageiro/análise , Fator de Transcrição TFIIDAssuntos
Proteínas de Ligação a DNA/genética , Mutação , Proteínas do Tecido Nervoso , Polidactilia/genética , Proteínas Repressoras , Fatores de Transcrição , Proteínas de Xenopus , Adulto , Sequência de Aminoácidos , Criança , Proteínas de Ligação a DNA/metabolismo , Face/anormalidades , Feminino , Hamartoma/diagnóstico , Humanos , Fatores de Transcrição Kruppel-Like , Imageamento por Ressonância Magnética , Masculino , Dados de Sequência Molecular , Linhagem , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Homologia de Sequência de Aminoácidos , Proteína Gli3 com Dedos de ZincoRESUMO
Seventy-two near full size long terminal repeats (LTRs) of human endogenous retrovirus of K-family (HERV-K) have been precisely located on the metric map of human chromosome 19. The LTR-related sequences were identified and assigned to cosmids by hybridization with two independent chromosome 19 specific cDNA clones corresponding to different parts of U3 region of LTR of HERV-K. The presence of full-size LTR sequences in a cosmid was further verified by PCR assay with a pair of primers complementary to the termini of the LTR. Coincidences of the LTR and the known genes positions are discussed.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 19/genética , Sequências Repetitivas de Ácido Nucleico/genética , Retroviridae/genética , Sequência de Bases , Cromossomos Humanos Par 19/virologia , Cosmídeos/genética , Genes/genética , Humanos , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
Greig cephalopolysyndactyly syndrome (GCPS, MIM 175700) is a rare autosomal dominant developmental disorder characterized by craniofacial abnormalities and post-axial and pre-axial polydactyly as well as syndactyly of hands and feet. Human GLI3, located on chromosome 7p13, is a candidate gene for the syndrome because it is interrupted by translocation breakpoints associated with GCPS. Since hemizygosity of 7p13 resulting in complete loss of one copy of GLI3 causes GCPS as well, haploinsufficiency of this gene was implicated as a mechanism to cause this developmental malformation. To determine if point mutations within GLI3 could be responsible for GCPS we describe the genomic sequences at the boundaries of the 15 exons and primer pair sequences for mutation analysis with polymerase chain reaction-based assays of the entire GLI3 coding sequences. In two GCPS cases, both of which did not exhibit obvious cytogenetic rearrangements, point mutations were identified in different domains of the protein, showing for the first time that Greig syndrome can be caused by GLI3 point mutations. In one case a nonsense mutation in exon X generates a stop codon truncating the protein in the C-H link of the first zinc finger. In the second case a missense mutation in exon XIV causes a Pro-->Ser replacement at a position that is conserved among GLI genes from several species altering a potential phosphorylation site.