Subject(s)
Abnormalities, Multiple/genetics , Heart Defects, Congenital/pathology , Noonan Syndrome/genetics , Protein Tyrosine Phosphatases/genetics , Abnormalities, Multiple/pathology , Adolescent , Adult , Child , Child, Preschool , DNA/chemistry , DNA/genetics , DNA Mutational Analysis , Developmental Disabilities/pathology , Family Health , Female , Genotype , Growth Disorders/pathology , Humans , Infant , Intracellular Signaling Peptides and Proteins , Lentigo/pathology , Male , Mutation , Noonan Syndrome/pathology , Pedigree , Phenotype , Polymorphism, Single-Stranded Conformational , Protein Tyrosine Phosphatase, Non-Receptor Type 11 , SyndromeABSTRACT
The ubiquitin fusion-degradation gene (UFD1L) encodes the human homologue of the yeast ubiquitin fusion-degradation 1 protein, an essential component of the ubiquitin-dependent proteolytic turnover and mRNA processing. Although the UFD1L gene has been mapped in the region commonly deleted in patients with DiGeorge syndrome (DGS)/velocardiofacial syndrome (VCFS), correlation between its haploinsufficiency and the phenotype has not yet been established. The only functional data available about mammalian Ufd1p is the ability to form a complex with the rat Npl4 protein, a component of the nuclear pore complex. In this paper we report the cloning and molecular characterization of the human NPL4 gene. This gene encodes for a protein 96% homologous to the rat Npl4, and 44 and 34% homologous to the C. elegans and S. cerevisiae Npl4 gene products, respectively. Fluorescence in situ hybridization experiments on human metaphases localized the NPL4 gene on the most telomeric region of chromosome 17q. Northern blots analysis on foetal and adult human tissues revealed a major approximately 4.5 kb transcript most abundant in heart, brain, kidney and skeletal muscle. In order to test a potential relationship between nuclear transport defects and some aspect of the DGS/VCFS phenotype, we also exclude the presence of mutations in the NPL4 coding sequence in a subset of patients with DGS/VCFS and no detectable 22q11 deletion or mutations at the UFD1L locus.
Subject(s)
Nuclear Pore Complex Proteins , Nuclear Proteins/genetics , Proteins/metabolism , Saccharomyces cerevisiae Proteins , Abnormalities, Multiple/genetics , Abnormalities, Multiple/pathology , Adaptor Proteins, Vesicular Transport , Amino Acid Sequence , Base Sequence , Blotting, Northern , Cell Cycle Proteins , Chromosome Banding , Chromosome Mapping , Chromosomes, Human, Pair 17/genetics , Cleft Palate/pathology , Cloning, Molecular , DNA, Complementary/genetics , DiGeorge Syndrome/genetics , Face/abnormalities , Female , Gene Expression , Gene Expression Regulation, Developmental , Heart Defects, Congenital/pathology , Humans , In Situ Hybridization, Fluorescence , Intracellular Signaling Peptides and Proteins , Molecular Sequence Data , Mutation , Nuclear Proteins/metabolism , Nucleocytoplasmic Transport Proteins , Polymorphism, Single Nucleotide/genetics , Protein Binding , Proteins/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Saccharomyces cerevisiae/genetics , Sequence Alignment , Sequence Homology, Amino Acid , Syndrome , Tissue Distribution , Two-Hybrid System TechniquesSubject(s)
Abnormalities, Multiple/genetics , Elastin/genetics , Abnormalities, Multiple/etiology , Abnormalities, Multiple/pathology , DNA/chemistry , DNA/genetics , DNA Mutational Analysis , Growth Disorders/pathology , Humans , Intellectual Disability/pathology , Polymorphism, Single-Stranded Conformational , SyndromeABSTRACT
Given q as the global frequency of the alleles causing a disease, any allele with a frequency higher than q minus the cumulative frequency of the previously known disease-causing mutations (threshold) cannot be the cause of that disease. This principle was applied to the analysis of cystic fibrosis transmembrane conductance regulator (CFTR) mutations in order to decide whether they are the cause of cystic fibrosis. A total of 191 DNA samples from random individuals from Italy, France, and Spain were investigated by DGGE (denaturing gradient gel electrophoresis) analysis of all the coding and proximal non-coding regions of the gene. The mutations detected by DGGE were identified by sequencing. The sample size was sufficient to select essentially all mutations with a frequency of at least 0.01. A total of 46 mutations was detected, 20 of which were missense mutations. Four new mutations were identified: 1341+28 C/T, 2082 C/T, L1096R, and I11131V. Thirteen mutations (125 G/C, 875+40 A/G, TTGAn, IVS8-6 5T, IVS8-6 9T, 1525-61 A/G, M470V, 2694 T/G, 3061-65 C/A, 4002 A/G, 4521 G/A, IVS8 TG10, IVS8 TG12) were classified as non-CF-causing alleles on the basis of their frequency. The remaining mutations have a cumulative frequency far exceeding q; therefore, most of them cannot be CF-causing mutations. This is the first random survey capable of detecting all the polymorphisms of the coding sequence of a gene.
