RESUMEN
Neuronal ceroid lipofuscinoses (NCLs) represent the most common group of inherited progressive encephalopathies in children. They are characterized by progressive loss of vision, mental and motor deterioration, epileptic seizures, and premature death. Rare adult forms of NCL with late onset are known as Kufs' disease. Loci underlying these adult forms remain unknown due to the small number of patients and genetic heterogeneity. Here we confirm that a late-onset form of NCL recessively segregates in US and French pedigrees of American Staffordshire Terrier (AST) dogs. Through combined association, linkage, and haplotype analyses, we mapped the disease locus to a single region of canine chromosome 9. We eventually identified a worldwide breed-specific variant in exon 2 of the Arylsulfatase G (ARSG) gene, which causes a p.R99H substitution in the vicinity of the catalytic domain of the enzyme. In transfected cells or leukocytes from affected dogs, the missense change leads to a 75% decrease in sulfatase activity, providing a functional confirmation that the variant might be the NCL-causing mutation. Our results uncover a protein involved in neuronal homeostasis, identify a family of candidate genes to be screened in patients with Kufs' disease, and suggest that a deficiency in sulfatase is part of the NCL pathogenesis.
Asunto(s)
Arilsulfatasas/genética , Enfermedades de los Perros/genética , Mutación Missense , Lipofuscinosis Ceroideas Neuronales/veterinaria , Transportadoras de Casetes de Unión a ATP/genética , Factores de Edad , Animales , Arilsulfatasas/deficiencia , Dominio Catalítico/genética , Línea Celular , Corteza Cerebelosa/metabolismo , Corteza Cerebelosa/patología , Corteza Cerebelosa/ultraestructura , Mapeo Cromosómico , Cromosomas de los Mamíferos/genética , Enfermedades de los Perros/enzimología , Perros , Femenino , Perfilación de la Expresión Génica , Frecuencia de los Genes , Genotipo , Haplotipos , Humanos , Masculino , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Linaje , Polimorfismo de Nucleótido Simple , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The conserved prpC, prkC, cpgA locus in Bacillus subtilis encodes respectively a Ser/Thr phosphatase, the cognate sensor kinase (containing an external PASTA domain suggested to bind peptidoglycan precursors) and CpgA, a small ribosome-associated GTPase that we have shown previously is implicated in shape determination and peptidoglycan deposition. In this study, in a search for targets of PrkC and PrpC, we showed that, in vitro, CpgA itself is phosphorylated on serine and threonine, and another GTPase, the translation factor EF-Tu, is also phosphorylated by the kinase on the conserved T384 residue. Both substrates are dephosphorylated by PrpC in vitro. In addition, we identified YezB, a 10.3 kDa polypeptide, and a component of the stressosome, as a substrate for both enzymes in vitro and apparently in vivo. We propose that the PrpC/PrkC/CpgA system constitutes an important element of a regulatory network involved in the coordination of cell wall expansion and growth in B. subtilis.
Asunto(s)
Bacillus subtilis/enzimología , Proteínas Bacterianas/metabolismo , Factor Tu de Elongación Peptídica/metabolismo , Fosfoproteínas Fosfatasas/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Bacillus subtilis/genética , Proteínas Bacterianas/genética , Escherichia coli/genética , Mutagénesis Sitio-Dirigida , Factor Tu de Elongación Peptídica/genética , Fosfoproteínas Fosfatasas/genética , Fosforilación , Proteínas Serina-Treonina Quinasas/genética , Serina/metabolismo , Especificidad por Sustrato , Treonina/metabolismoRESUMEN
Dogs differ greatly in their morphological characteristics including various tail phenotypes. Congenitally short-tailed dogs are present in many breeds; however, the causative mutation located in the T-box transcription factor T gene (C189G) had only been described in the bobtailed Pembroke Welsh Corgis. We investigated here the presence of the T gene mutation in 23 other breeds (360 dogs, including 156 natural short tailed) in which natural bobtailed dogs exist. In the 17 breeds in which the C189G mutation was observed, there was a perfect correlation between this mutation and the short-tail phenotype. However, 6 breeds did not carry the known substitution or any other mutations in the T gene coding regions. No dogs were found to be homozygous for the C189G mutation, suggesting that the homozygous condition is lethal. In order to study the effect of the T gene mutation on litter size, we compared the number of puppies born from short-tailed parents to that born from long-tailed parents. In the Swedish Vallhund breed, we observed a 29% decrease in the litter size when both parents were short tailed. Given that the T gene mutation is not present in all breeds of short-tailed dog, there must be yet other genetic factors affecting tail phenotypes to be discovered.
Asunto(s)
Cruzamiento , Mutación , Proteínas de Dominio T Box/genética , Cola (estructura animal)/anatomía & histología , Animales , Cruzamientos Genéticos , Perros , Evolución Molecular , Femenino , Tamaño de la Camada/genética , Mutación/fisiología , Fenotipo , Filogenia , EmbarazoRESUMEN
BACKGROUND: Several forms of progressive retinal atrophy (PRA) segregate in more than 100 breeds of dog with each PRA segregating in one or a few breeds. This breed specificity may be accounted for by founder effects and genetic drift, which have reduced the genetic heterogeneity of each breed, thereby facilitating the identification of causal mutations. We report here a new form of PRA segregating in the Border Collie breed. The clinical signs, including the loss of night vision and a progressive loss of day vision, resulting in complete blindness, occur at the age of three to four years and may be detected earlier through systematic ocular fundus examination and electroretinography (ERG). RESULTS: Ophthalmic examinations performed on 487 dogs showed that affected dogs present a classical form of PRA. Of those, 274 have been sampled for DNA extraction and 87 could be connected through a large pedigree. Segregation analysis suggested an X-linked mode of transmission; therefore both XLPRA1 and XLPRA2 mutations were excluded through the genetic tests. CONCLUSION: Having excluded these mutations, we suggest that this PRA segregating in Border Collie is a new XLPRA (XLPRA3) and propose it as a potential model for the homologous human disease, X-Linked Retinitis Pigmentosa.