A novel familial mutation in the PCSK1 gene that alters the oxyanion hole residue of proprotein convertase 1/3 and impairs its enzymatic activity.

Four siblings presented with congenital diarrhea and various endocrinopathies. Exome sequencing and homozygosity mapping identified five regions, comprising 337 protein-coding genes that were shared by three affected siblings. Exome sequencing identified a novel homozygous N309K mutation in the proprotein convertase subtilisin/kexin type 1 (PCSK1) gene, encoding the neuroendocrine convertase 1 precursor (PC1/3) which was recently reported as a cause of Congenital Diarrhea Disorder (CDD). The PCSK1 mutation affected the oxyanion hole transition state-stabilizing amino acid within the active site, which is critical for appropriate proprotein maturation and enzyme activity. Unexpectedly, the N309K mutant protein exhibited normal, though slowed, prodomain removal and was secreted from both HEK293 and Neuro2A cells. However, the secreted enzyme showed no catalytic activity, and was not processed into the 66 kDa form. We conclude that the N309K enzyme is able to cleave its own propeptide but is catalytically inert against in trans substrates, and that this variant accounts for the enteric and systemic endocrinopathies seen in this large consanguineous kindred

Установление границ охранной зоны линейного сооружения – магистральный газопровод "НГПЗ - Парабель"

Составлено графическое описание местоположения границ зон с особыми условиями использования территорий границ охранной зоны линейного сооружения – магистральный газопровод "НГПЗ - Парабель".A graphic description of the location of the boundaries of the zones with special conditions for the use of the territories of the boundaries of the protection zone of the linear structure – "the NGPZ-Parabel" gas pipeline has been compiled

Applications of Next-Generation DNA Sequencing to the Identification of Rare Variants in Congenital Disorders of the Intestine and Brain

High throughput, massively parallel DNA sequencing provides a powerful technology to study the human genome and to identify variations in DNA that cause disease. Sequencing the protein coding region of the genome (`whole-exome sequencing') is a cost effective method to search the part of the genome that is most likely to harbor disease related mutations.We developed software methods to process sequencing data and to annotate variants with data on genes, function, conservation, expression, diseases, pathways, and protein structure. We applied whole-exome sequencing to search for the molecular basis of disease in three projects: 1) a cohort of patients with congenital diarrheal disorders (CDDs); 2) a cohort of patients with congenital chronic intestinal pseudo-obstruction (CIPO) or the related disease, megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIH); and 3) four siblings with infantile pontocerebellar hypoplasia and spinal motor neuron degeneration.We sequenced 45 probands from diverse ethnic backgrounds who were diagnosed with a variety of CDDs of probable, but unknown genetic cause. Patients had been diagnosed with generalized malabsorptive diarrhea, selective nutrient malabsorption, secretory diarrhea, and infantile IBD. We found homozygous or compound heterozygous mutations, 25 of them novel, in genes known to be associated with CDDs in 27 cases (60%). The genes implicated were ADAM17, DGAT1, EPCAM, IL10RA, MALT1, MYO5B, NEUROG3, PCSK1, SI, SKIV2L, SLC26A3, and SLC5A.With whole-exome sequencing in a cohort of 20 patients with congenital CIPO or MMIH, we identified a subset of 10 cases with potentially damaging de-novo dominant acting mutations at highly conserved loci in the ACTG2 gene, encoding actin, gamma-enteric smooth muscle precursor, a protein essential to the functioning of muscle cells in the intestinal wall.By exome sequencing, we discovered rare recessive mutations in EXOSC3 (encoding exosome component 3) that were responsible for pontocerebellar hypoplasia and spinal motor neuron degeneration in the four probands, and identified identical and additional novel mutations in a large percentage of other children with the same disorder.In conclusion, we demonstrated that whole-exome sequencing is an effective approach for the identification of casual mutations in that may escape detection with standard practice involving a complex diagnostic workup and targeted gene sequencing

Rich annotation of DNA sequencing variants by leveraging the Ensembl Variant Effect Predictor with plugins

High-throughput DNA sequencing has become a mainstay for the discovery of genomic variants that may cause disease or affect phenotype. A next-generation sequencing pipeline typically identifies thousands of variants in each sample. A particular challenge is the annotation of each variant in a way that is useful to downstream consumers of the data, such as clinical sequencing centers or researchers. These users may require that all data storage and analysis remain on secure local servers to protect patient confidentiality or intellectual property, may have unique and changing needs to draw on a variety of annotation data sets and may prefer not to rely on closed-source applications beyond their control. Here we describe scalable methods for using the plugin capability of the Ensembl Variant Effect Predictor to enrich its basic set of variant annotations with additional data on genes, function, conservation, expression, diseases, pathways and protein structure, and describe an extensible framework for easily adding additional custom data sets

A novel familial mutation in the PCSK1 gene that alters the oxyanion hole residue of proprotein convertase 1/3 and impairs its enzymatic activity.

Four siblings presented with congenital diarrhea and various endocrinopathies. Exome sequencing and homozygosity mapping identified five regions, comprising 337 protein-coding genes that were shared by three affected siblings. Exome sequencing identified a novel homozygous N309K mutation in the proprotein convertase subtilisin/kexin type 1 (PCSK1) gene, encoding the neuroendocrine convertase 1 precursor (PC1/3) which was recently reported as a cause of Congenital Diarrhea Disorder (CDD). The PCSK1 mutation affected the oxyanion hole transition state-stabilizing amino acid within the active site, which is critical for appropriate proprotein maturation and enzyme activity. Unexpectedly, the N309K mutant protein exhibited normal, though slowed, prodomain removal and was secreted from both HEK293 and Neuro2A cells. However, the secreted enzyme showed no catalytic activity, and was not processed into the 66 kDa form. We conclude that the N309K enzyme is able to cleave its own propeptide but is catalytically inert against in trans substrates, and that this variant accounts for the enteric and systemic endocrinopathies seen in this large consanguineous kindred