acid-phosphatase and Chromosome-Deletion

Only a few genetic causes for childhood obesity have been identified to date. Copy number variants (CNVs) are known to contribute to obesity, both syndromic (15q11.2 deletions, Prader-Willi syndrome) and nonsyndromic (16p11.2 deletions) obesity.. To study the contribution of CNVs to early-onset obesity and evaluate the expression of candidate genes in subcutaneous adipose tissue.. A case-control study in a tertiary academic center.. CNV analysis was performed on 90 subjects with early-onset obesity and 67 normal-weight controls. Subcutaneous adipose tissue from body mass index-discordant siblings was used for the gene expression analyses.. We used custom high-density array comparative genomic hybridization with exon resolution in 1989 genes, including all known obesity loci. The expression of candidate genes was assessed using microarray analysis of messenger RNA from subcutaneous adipose tissue.. We identified rare CNVs in 17 subjects (19%) with obesity and 2 controls (3%). In three cases (3%), the identified variant involved a known syndromic lesion (22q11.21 duplication, 1q21.1 deletion, and 16p11.2 deletion, respectively), although the others were not known. Seven CNVs in 10 families were inherited and segregated with obesity. Expression analysis of 37 candidate genes showed discordant expression for 10 genes (PCM1, EFEMP1, MAMLD1, ACP6, BAZ2B, SORBS1, KLF15, MACROD2, ATR, and MBD5).. Rare CNVs contribute possibly pathogenic alleles to a substantial fraction of children with early-onset obesity. The involved genes might provide insights into pathogenic mechanisms and involved cellular pathways. These findings highlight the importance of CNV screening in children with early-onset obesity.

Topics: Abnormalities, Multiple; Acid Phosphatase; Adolescent; Adult; Ataxia Telangiectasia Mutated Proteins; Autistic Disorder; Autoantigens; Case-Control Studies; Cell Cycle Proteins; Child; Child, Preschool; Chromosome Deletion; Chromosome Disorders; Chromosome Duplication; Chromosomes, Human, Pair 1; Chromosomes, Human, Pair 16; Chromosomes, Human, Pair 22; Comparative Genomic Hybridization; DiGeorge Syndrome; DNA Copy Number Variations; DNA Repair Enzymes; DNA-Binding Proteins; Extracellular Matrix Proteins; Female; Humans; Hydrolases; Intellectual Disability; Kruppel-Like Transcription Factors; Male; Megalencephaly; Microfilament Proteins; Nuclear Proteins; Pediatric Obesity; Proteins; RNA, Messenger; Siblings; Subcutaneous Fat; Transcription Factors; Transcription Factors, General; Transcriptome; Young Adult

The genetic basis of the varying ability to reduce nitrate in strains belonging to different biovars and subspecies of plague-causing microbe has been investigated and the inability to reduce nitrate observed in different intraspecies groups of Yersinia pestis has been shown to stem from mutations in different genes involved in the expression of this trait. The absence of denitrifying activity in strains of altaica and hissarica subspecies was not due to a mutation at position 613 of the periplasmic reductase napA observed in the strains of the biovar medievalis of the main subspecies, but rather was due to a mutation in the sequence encoding the nitrate-binding domain of the ABC transporter protein SsuA; a thymine insertion (+T) was detected at position 302 from the start of the ssuA gene. Five strains of biovar antiqua isolated at different times in Mongolia, China, and Africa were shown to lack the ability to reduce nitrate. A PCR test targeting two chromosomal regions containing deletions of 19 and 24 bp in size has been developed for the identification of strains of the biovar medievalis. This test can be combined with the test for the marker mutation in the napA gene for a more reliable detection of Y. pestis strains belonging to this biovar.

Topics: Acid Phosphatase; ATP-Binding Cassette Transporters; Chromosome Deletion; Mutation; Nitrates; Periplasmic Proteins; Yersinia pestis

The PHO80 and PHO85 gene products encode proteins necessary for the repression of transcription from the major acid phosphatase gene (PHO5) of Saccharomyces cerevisiae. The deduced amino acid sequences of these genes have revealed that PHO85 is likely to encode a protein kinase, whereas no potential function has been revealed for PHO80. We undertook several approaches to aid in the elucidation of the PHO80 function, including deletion analysis, chemical mutagenesis, and expression analysis. DNA deletion analysis revealed that residues from both the carboxy- and amino-terminal regions of the protein, amounting to a total of 21% of the PHO80 protein, were not required for function with respect to repressor activity. Also, 10 independent single-amino-acid changes within PHO80 which resulted in the failure to repress PHO5 transcription were isolated. Nine of the 10 missense mutations resided in two subregions of the PHO80 molecule. In addition, expression analysis of the PHO80 and PHO85 genes suggested that the PHO85 gene product was not necessary for PHO80 expression and that the PHO85 gene was expressed at much higher levels in the cell than was the PHO80 gene. Furthermore, high levels of PHO80 were shown to suppress the effect of a PHO85 deletion at a level close to full repression. Implications for the function of the negative regulators in this system are discussed.

Topics: Acid Phosphatase; beta-Galactosidase; Chromosome Deletion; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Genes, Fungal; Genes, Regulator; Genotype; Kinetics; Mutagenesis, Site-Directed; Saccharomyces cerevisiae; Transcription, Genetic

We have studied patients with Duchenne muscular dystrophy (DMD), DMD together with glycerol kinase (GK) deficiency, or DMD together with both GK deficiency and congenital adrenal hypoplasia (AHC). Analysis of deletions in these patients allows the mapping of these mutations in Xp21. The following order is proposed: Xpter - L1 - AHC - GK - DMD - Xcen. One of the boys with DMD, GK, and AHC is shown by pulsed-field-gel electrophoresis to have a deletion which has a proximal endpoint at least 500 kb distal from the pERT87 (DXS164) locus.

Topics: Acid Phosphatase; Adrenal Insufficiency; Cell Line; Child; Child, Preschool; Chromosome Deletion; Chromosome Mapping; DNA; Glycerol Kinase; Humans; Male; Phosphotransferases; Sex Chromosome Aberrations; X Chromosome

The repressible Saccharomyces cerevisiae acid phosphatase (APase) coded by the PHO5 gene is a cell wall glycoprotein that follows the yeast secretory pathway. We used in vitro mutagenesis to construct a deletion (delta SP) including the entire signal sequence and four amino acids of the mature sequence of APase. An APase-deficient yeast strain was transformed with a high-copy-number plasmid carrying the PHO5/delta SP gene. When expressed in vivo, the PHO5/delta SP gene product accumulated predominantly as an inactive, unglycosylated form located inside the cell. A large part of this unglycosylated precursor underwent proteolytic degradation, but up to 30% of it was translocated, core glycosylated, and matured by the addition of mannose residues, before reaching the cell wall. It appears, therefore, that the signal sequence is important for efficient translocation and core glycosylation of yeast APase but that it is not absolutely necessary for entry of the protein into the yeast secretory pathway. mRNA obtained by in vitro transcription of PHO5 and PHO5/delta SP genes were translated in vitro in the presence of either reticulocyte lysate and dog pancreatic microsomes or yeast lysate and yeast microsomes. The PHO5 gene product was translocated and core glycosylated in the heterologous system and less efficiently in the homologous system. We were not able to detect any translocation or glycosylation of PHO5/delta SP gene product in the heterologous system, but a very small amount of core suppression of glycosylated material could be evidenced in the homologous system.

Topics: Acid Phosphatase; Amino Acid Sequence; Animals; Biological Transport; Chromosome Deletion; Dogs; Glycoproteins; Immunologic Techniques; In Vitro Techniques; Kinetics; Microsomes; Molecular Weight; Protein Processing, Post-Translational; Protein Sorting Signals; Saccharomyces cerevisiae; Structure-Activity Relationship

We report a patient with a de novo interstitial deletion of the short arm of chromosome 2 (p23p25). The patient had microcephaly with prominent forehead and occiput, narrow rectangular face, clinodactyly, failure to thrive, delayed psychomotor development, and seizures. Maternal serum alpha-fetoprotein was undetectable at 18 weeks of gestation. Heterozygosity at the red cell acid phosphatase locus (SRO-2p25) and normal levels of red cell malate dehydrogenase (SRO-2p23) are findings consistent with the presence of genetic material from bands 2p25 and 2p23.

Topics: Abnormalities, Multiple; Acid Phosphatase; Chromosome Aberrations; Chromosome Deletion; Chromosome Disorders; Chromosomes, Human, Pair 2; Erythrocytes; Genetic Markers; Humans; Infant, Newborn; Intellectual Disability; Malate Dehydrogenase; Male; Microcephaly

The locus for acid phosphatase (ACP1) had been alternately assigned to two conflicting regions on the short arm of chromosome 2. We present a clinical and cytogenetic report of one patient who has an interstitial deletion of 2, del(2) (p23p25.1), and a cytogenetic study of another cell line with an interstitial deletion of 2p (p23.1p25.1). Because both patients are heterozygotes for ACP1, the assignment of ACP1 to 2p25.1----pter is supported.

Topics: Abnormalities, Multiple; Acid Phosphatase; Child; Chromosome Aberrations; Chromosome Deletion; Chromosome Disorders; Chromosome Mapping; Chromosomes, Human, Pair 2; Genetic Markers; Humans; Infant, Newborn; Intellectual Disability; Microcephaly; Seizures

It was found that both poor selection pressure and a variable rate of plasmid loss were present in the system studied and that both have significant effects on continuous reactor operation. At least some of these effects were analyzed by a simple model. At this point, experimental analysis for extracellular levels of tryptophan sufficient to support X- growth (1-4 mg/l) has given contradictory results. This has at least partially indicated the effect may be an intracellular one, and thus the culture history would be critical in such experiments. Since the system studied is not atypical of recombinant cultures, it leads one to speculate on the generality of the phenomena and its extent in other cultures. If important, the use of double auxotrophs or auxotrophs that are mutant in a metabolite for which the cell has a greater growth requirement should be used. Additionally, the presence of higher copy numbers in yeast at lower growth rates also leads one to speculate on how these apparently contradictory phenomena are related.

Topics: Acid Phosphatase; Chromosome Deletion; DNA, Recombinant; Fermentation; Genes; Genes, Fungal; Kinetics; Plasmids; Recombinant Proteins; Recombination, Genetic; Saccharomyces cerevisiae

The expression of the PHO5 gene of Saccharomyces cerevisiae is transcriptionally regulated in response to the level of inorganic phosphate present in the growth medium. We have identified, by DNA deletion analysis, the sequences (upstream activator sequences) that mediate this response. The sequence 5' CTGCACAAATG 3' is present in two copies located within a 60-base-pair region. The presence of a single copy of the sequence is sufficient for the phosphate-mediated transcriptional response. In addition, a DNA fragment that contains two copies of this sequence will act to repress transcription of a CYC1-lacZ fusion when placed either upstream or downstream of the CYC1 activator sequence.

Topics: Acid Phosphatase; Base Sequence; Chromosome Deletion; Cloning, Molecular; Enzyme Repression; Genes; Genes, Fungal; Genes, Regulator; Phosphates; Plasmids; Saccharomyces cerevisiae; Transcription, Genetic

We studied ultrastructural localization of acid phosphatase in derepressed Saccharomyces cerevisiae cells transformed with a multicopy plasmid carrying either the wild-type PHO5 gene or a PHO5 gene deleted in the region overlapping the signal peptidase cleavage site. Wild-type enzyme was located in the cell wall, as was 50% of the modified protein, which carried high-mannose-sugar chains. The remaining 50% of the protein was active and core glycosylated, and it accumulated in the endoplasmic reticulum cisternae. The signal peptide remained uncleaved in both forms. Cells expressing the modified protein exhibited an exaggerated endoplasmic reticulum with dilated lumen.

Topics: Acid Phosphatase; Chromosome Deletion; Endopeptidases; Endoplasmic Reticulum; Genes; Genes, Fungal; Membrane Proteins; Microscopy, Electron; Mutation; Protein Sorting Signals; Saccharomyces cerevisiae; Serine Endopeptidases

We transformed Saccharomyces cerevisiae with a high-copy-number plasmid carrying either the wild-type gene coding for a repressible cell surface acid phosphatase or two modified genes whose products lack a 13- or 14-amino-acid segment spanning or immediately adjacent to the signal peptidase cleavage site. The wild-type gene product underwent proteolytic cleavage of the signal peptide, core glycosylation, and outer chain glycosylation. The deletion spanning the signal peptidase cleavage site led to an unprocessed protein. This modified protein exhibited core glycosylation, whereas its outer chain glycosylation was severely inhibited. Secretion of the deleted protein was impaired, and active enzyme accumulated within the cell. The deletion immediately adjacent to the signal peptidase cleavage site exhibited only a small decrease in the efficiency of processing and had no effect on the efficiency of secretion.

Topics: Acid Phosphatase; Amino Acid Sequence; Base Sequence; Binding Sites; Carbohydrate Metabolism; Chromosome Deletion; DNA Restriction Enzymes; Endopeptidases; Membrane Proteins; Plasmids; Protein Biosynthesis; Saccharomyces cerevisiae; Serine Endopeptidases

Maximum likelihood analysis for linkage between autosomal dominant aniridia and 12 biochemical and serological markers in a single large family showed a probable linkage between autosomal dominant aniridia and the enzyme acid phosphatase-1. The presence of an autosomal dominant aniridia gene linked to acid phosphatase-1 on chromosome arm 2p and the existence of an aniridia syndrome resulting from deletion of band 13 of the short arm of chromosome 11 establishes a chromosome basis for genetic heterogeneity of aniridia phenotypes.

Topics: Acid Phosphatase; Chromosome Aberrations; Chromosome Deletion; Chromosome Disorders; Chromosomes, Human, 1-3; Chromosomes, Human, 4-5; Computers; Congenital Abnormalities; Genes, Dominant; Genetic Linkage; Humans; Iris; Pedigree; Polymorphism, Genetic

The human red cell acid phosphatase (ACP1) locus was assigned to region 2p23 leads to 2pter by Ferguson-Smith et al [3], more specifically to 2p23 by Hamerton et al [5]. We describe two unrelated patients with deletion of chromosome 2, with similar breakpoints in the distal portion of band p23 (del(2) (p23)). ACP1 typing in both patients revealed heterozygous BA phenotypes. Thus, we assign the locus for ACP1 to the distal portion of 2p23.

Topics: Abnormalities, Multiple; Acid Phosphatase; Chromosome Deletion; Chromosome Mapping; Chromosomes, Human, 1-3; Female; Genetic Markers; Humans; Infant, Newborn; Male; Phenotype

Topics: Acid Phosphatase; Animals; Chromosome Deletion; Chromosome Mapping; Drosophila melanogaster; Female; Frozen Sections; Ganglia; Genetic Linkage; Genotype; Glucosephosphate Dehydrogenase; Histocytochemistry; Male; Mosaicism; Nervous System; Phosphogluconate Dehydrogenase; Sex Chromosomes; Translocation, Genetic

Topics: Acid Phosphatase; Blood Group Antigens; Chromosome Aberrations; Chromosome Deletion; Chromosomes, Human, 1-3; Erythrocytes; Female; Genetic Linkage; Humans; Male

Analysis of human-Chinese hamster somatic cell hybrids with spontaneously derived chromosome structural changes has provided data for the regional and subregional localization of gene loci which have previously been assigned to human chromosomes 2, 12, and X. Correlation of the expression of human gene loci with the human chromosome complements present in somatic cell hybrids indicates that the cytoplasmic malate dehydrogenase (MDH1) locus is in the 2p23yields2pter region, and red cell acid phosphatase (AcP1) is at or adjacent to 2p23. The cytoplasmic isocitrate dehydrogenase (IDH1) locus is at or adjacent to 2q11, peptidase B (Pep B) is at or adjacent to 12q21, lactate dehydrogenase B (LDH B) is in the 12q21yiedls12pter region, glucose-6-phosphate dehydrogenase (G6PD) is in the Xq24yieldsXqter region, and the gene loci for phosphoglycerate kinase (PGK), alpha-galactosidase (alpha-gal), and hypoxanthine guanine phosphoribosyltransferase (GPRT) are in the Xp21yieldsXq24 region.

Topics: Acid Phosphatase; Animals; Carbohydrate Dehydrogenases; Chromosome Aberrations; Chromosome Deletion; Chromosome Mapping; Chromosomes, Human, 1-3; Chromosomes, Human, 6-12 and X; Cricetinae; Genes; Genetic Linkage; Glucosephosphate Dehydrogenase; Humans; Hybrid Cells; Isocitrate Dehydrogenase; Karyotyping; Malate Dehydrogenase; Peptide Hydrolases; Sex Chromosomes; Translocation, Genetic