RESUMO
Schwartz-Jampel syndrome (SJS1) is a rare autosomal recessive disorder characterized by permanent myotonia (prolonged failure of muscle relaxation) and skeletal dysplasia, resulting in reduced stature, kyphoscoliosis, bowing of the diaphyses and irregular epiphyses. Electromyographic investigations reveal repetitive muscle discharges, which may originate from both neurogenic and myogenic alterations. We previously localized the SJS1 locus to chromosome 1p34-p36.1 and found no evidence of genetic heterogeneity. Here we describe mutations, including missense and splicing mutations, of the gene encoding perlecan (HSPG2) in three SJS1 families. In so doing, we have identified the first human mutations in HSPG2, which underscore the importance of perlecan not only in maintaining cartilage integrity but also in regulating muscle excitability.
Assuntos
Proteoglicanas de Heparan Sulfato/genética , Mutação , Osteocondrodisplasias/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Análise Mutacional de DNA , Primers do DNA/genética , Feminino , Proteoglicanas de Heparan Sulfato/química , Humanos , Masculino , Camundongos , Dados de Sequência Molecular , Linhagem , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , Especificidade da EspécieRESUMO
Autosomal dominant hereditary spastic paraplegia (AD-HSP) is a genetically heterogeneous neurodegenerative disorder characterized by progressive spasticity of the lower limbs. Among the four loci causing AD-HSP identified so far, the SPG4 locus at chromosome 2p2-1p22 has been shown to account for 40-50% of all AD-HSP families. Using a positional cloning strategy based on obtaining sequence of the entire SPG4 interval, we identified a candidate gene encoding a new member of the AAA protein family, which we named spastin. Sequence analysis of this gene in seven SPG4-linked pedigrees revealed several DNA modifications, including missense, nonsense and splice-site mutations. Both SPG4 and its mouse orthologue were shown to be expressed early and ubiquitously in fetal and adult tissues. The sequence homologies and putative subcellular localization of spastin suggest that this ATPase is involved in the assembly or function of nuclear protein complexes.
Assuntos
Adenosina Trifosfatases/genética , Mutação , Paraplegia Espástica Hereditária/genética , Adenosina Trifosfatases/química , Adenosina Trifosfatases/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Sequência de Bases , Células Cultivadas , Clonagem Molecular , Análise Mutacional de DNA , Éxons/genética , Etiquetas de Sequências Expressas , Humanos , Íntrons/genética , Camundongos , Mitocôndrias Musculares/metabolismo , Dados de Sequência Molecular , Fosforilação Oxidativa , RNA Mensageiro/análise , RNA Mensageiro/genética , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Paraplegia Espástica Hereditária/enzimologia , Paraplegia Espástica Hereditária/metabolismo , Paraplegia Espástica Hereditária/patologia , EspastinaRESUMO
Schwartz-Jampel syndrome (SJS) is a rare autosomal recessive disorder characterized by the association of myotonia with chondrodysplasia. A positional cloning strategy allowed the localization and the identification of perlecan as the disease causing gene. Another human recessive disorder, the Dyssegmental Dysplasia, Silverman-Handmaker type (DDSH), is caused by functional null mutations of the perlecan gene. A gene-dosage effect seems to account for the correlation between the phenotype and the mutations within the gene: SJS would be associated with hypomorph mutations of the perlecan gene and DDSH would be due to the absence of functional perlecan. Perlecan is the major heparan sulfate proteoglycan of extracellular matrix and basement membranes that displays various functions. Based on these functions, several hypotheses are evoked to explain chondrodysplasia and the unusual myotonia in SJS. Mouse models are invaluable tools to explore these hypotheses. Since knock-out mice develop a phenotype similar to DDSH, we have developed a SJS mouse model by reproducing the hypomorph effect of SJS perlecan mutation in this species. The characterization of this mouse model will help to understand the pathophysiological mechanism leading to this multisystemic human disorder.
Assuntos
Proteoglicanas de Heparan Sulfato/deficiência , Osteocondrodisplasias/genética , Animais , Membrana Basal/metabolismo , Cartilagem/metabolismo , Modelos Animais de Doenças , Fibroblastos/metabolismo , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/fisiologia , Humanos , Camundongos , Camundongos Knockout , Mutação de Sentido Incorreto , Osteocondrodisplasias/metabolismo , Osteocondrodisplasias/fisiopatologiaRESUMO
Autosomal dominant familial spastic paraplegia (AD-FSP) is a genetically heterogeneous, neurodegenerative disorder characterized by spasticity and progressive weakness in the lower limbs. Anticipation has been suggested to occur and an association between expanded CAG/CTG repeats and AD-FSP linked to the SPG4 locus (2p21-p24) has been described. In this study, 42 affected individuals from six SPG4 families were screened for expanded CAG/CTG repeats using the repeat expansion detection (RED) method. Large RED products (range 180-240 nucleotides) corresponding in size to repeats at the ERDA1 locus were detected in eight patients and at the CTG 18.1 locus in one patient. The large ERDA1 repeats did not segregate with the disorder within families. Mean age at onset and index of severity were not significantly different between patients with or without expanded RED products. Furthermore, no abnormal proteins were found by Western blot in 15 selected patient samples as compared with controls, using the 1C2 antibody, which detects long polyglutamine stretches. Thus, in contrast to previous reports, our study provides evidence against the hypothesis that a large translated CAG repeat expansion is the basis of SPG4. We propose that mechanisms other than large pathogenic CAG/CTG repeats may account for the disease in the SPG4 families tested here.
Assuntos
Cromossomos Humanos Par 2/genética , Paraplegia/genética , Repetições de Trinucleotídeos/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Ligação Genética/genética , Humanos , Pessoa de Meia-IdadeRESUMO
Hereditary spastic paraplegia (HSP) comprises a group of clinically and genetically heterogeneous disorders causing progressive spasticity and weakness of the lower limbs. We report a large family of French descent with autosomal dominant pure HSP. We excluded genetic linkage to the known loci causing HSP and performed a genomewide search. We found evidence for linkage of the disorder to polymorphic markers on chromosome 2q24-q34: a maximum LOD score of 3. 03 was obtained for marker D2S2318. By comparison with families having linkage to the major locus of pure autosomal dominant HSP (SPG4 on chromosome 2p), there were significantly more patients without Babinski signs, with increased reflexes in the upper limbs, and with severe functional handicaps.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 2/genética , Genes Dominantes/genética , Paraplegia/genética , Adulto , Alelos , Feminino , França/etnologia , Haplótipos/genética , Humanos , Escore Lod , Masculino , Paraplegia/fisiopatologia , Linhagem , Fenótipo , Polimorfismo Genético/genéticaRESUMO
Autosomal dominant hereditary spastic paraplegia (AD-HSP) is a genetically heterogeneous disorder characterized by progressive spasticity of the lower limbs. A major locus (SPG4) causing AD-HSP in about 40% of the families was mapped to chromosome 2p. The analysis of six SPG4-linked AD-HSP families using the RED procedure previously showed the expansion of a CAG repeat in affected individuals. To identify the gene responsible for this form of HSP, we have constructed a 3.5-Mb YAC contig flanked by loci D2S400 and D2S367, have subcloned five of these YACs spanning the candidate region into cosmids, and screened these cosmid libraries for the presence of CAG repeat sequences. Four CAG repeats have been identified but none of them is expanded in 26 patients from 13 SPG4-linked AD-HSP families. A gene map comprising 21 transcripts was established using expressed sequence tags (ESTs) assigned previously to this region of 2p21-p22 with radiation hybrid panels GeneBridge 4 and G3. Full-length cDNAs corresponding to the 14 ESTs mapping to the SPG4 interval flanked by loci D2S352 and D2S2347 were isolated and sequenced. None contains a CAG repeat in its coding sequence. Finally, we have assembled a BAC contig composed of 37 clones that were also screened for the presence of CAG repeats; this failed to detect additional repeats to those identified on YACs.
Assuntos
Cromossomos Humanos Par 2/genética , Paraplegia Espástica Hereditária/genética , Repetições de Trinucleotídeos/genética , Cromossomos Bacterianos/genética , Clonagem Molecular , Mapeamento de Sequências Contíguas , Etiquetas de Sequências Expressas , Humanos , Repetições de Microssatélites , Análise de Sequência de DNARESUMO
Autosomal dominant familial spastic paraplegia (AD-FSP) is a genetically heterogeneous neurodegenerative disorder characterized by progressive spasticity of the lower limbs. Three loci on chromosome 14q (SPG3), 2p (SPG4), and 15q (SPG6) were shown to be responsible for AD-FSP. Analysis of recombination events in three SPG3-linked families allowed us to narrow the critical interval from 9 to 5 cM. An approximately 5-Mb YAC contig comprising 32 clones and 90 STSs was built from D14S301 to D14S991, encompassing this region of 14q21. Fifty-six ESTs assigned previously to this region with radiation hybrid (RH) panels Genebridge 4 and G3 were precisely localized on the YAC contig. The 90 STSs positioned on the contig were tested on the TNG RH panel to compare our YAC-based map with an RH map at a high level of resolution. Comparison between our map and the whole genome mapping data on this interval of chromosome 14q is discussed.
Assuntos
Cromossomos Humanos Par 14/genética , Genoma Humano , Paraplegia Espástica Hereditária/genética , Mapeamento Cromossômico , Mapeamento de Sequências Contíguas , Etiquetas de Sequências Expressas , Saúde da Família , Feminino , Humanos , Células Híbridas/efeitos da radiação , Masculino , Repetições de Microssatélites , Linhagem , Sitios de Sequências Rotuladas , Transcrição GênicaRESUMO
Autosomal dominant hereditary spastic paraplegia (AD-HSP) is a group of genetically heterogeneous neurodegenerative disorders characterized by pro- gressive spasticity of the lower limbs. Five AD-HSP loci have been mapped to chromosomes 14q, 2p, 15q, 8q and 12q. The SPG4 locus at 2p21-p22 has been shown to account for approximately 40% of all AD-HSP families. SPG4 encoding spastin, a putative nuclear AAA protein, has recently been identified. Here, sequence analysis of the 17 exons of SPG4 in 87 unrelated AD-HSP patients has resulted in the detection of 34 novel mutations. These SPG4 mutations are scattered along the coding region of the gene and include all types of DNA modification including missense (28%), nonsense (15%) and splice site point (26.5%) mutations as well as deletions (23%) and insertions (7.5%). The clinical analysis of the 238 mutation carriers revealed a high proportion of both asymptomatic carriers (14/238) and patients unaware of symptoms (45/238), and permitted the redefinition of this frequent form of AD-HSP.
Assuntos
Adenosina Trifosfatases/genética , Genes Dominantes , Mutação , Paraplegia/genética , Adenosina Trifosfatases/fisiologia , Adolescente , Adulto , Idoso , Criança , Códon sem Sentido , Genótipo , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Fenótipo , Polimorfismo Genético , Splicing de RNA , EspastinaRESUMO
We report the clinical features of 12 families with autosomal dominant spastic paraplegia (ADSP) linked to the SPG4 locus on chromosome 2p, the major locus for this disorder that accounts for approximately 40% of the families. Among 93 gene carriers, 32 (34%) were unaware of symptoms but were clinically affected. Haplotype reconstruction showed that 90% of the asymptomatic gene carriers presented increased reflexes and/or extensor plantar responses independent of age at examination. The mean age at onset was 29 years, ranging from 1 to 63 years. Intra- as well as inter-familial variability of age at onset was important, but did not result from anticipation. Phenotype-genotype correlations and comparison with SPG3 and SPG5 families indicated that despite the variability of age at onset, SPG4 is a single genetic entity but no clinical features distinguish individual SPG4 patients from those with SPG3 or SPG5 mutations.