DOCSLIB.ORG

Regulation of Hypoxanthine DNA Glycosylase in Normal Human and Bloom's Syndrome Fibroblasts1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Regulation of Hypoxanthine DNA Glycosylase in Normal Human and Bloom's Syndrome Fibroblasts1 [CANCER RESEARCH 46, 3756-3761, August 1986] Regulation of Hypoxanthine DNA Glycosylase in Normal Human and Bloom's Syndrome Fibroblasts1 Philip Dehazya2 and Michael A. Sirover3 Fels Research institute and the Department of Pharmacology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140 ABSTRACT cells appears to be dependent on the proliferative state of the cell. Previous studies described the relationship between the The regulation of the base excision repair enzyme hypoxanthine DNA increase in the activity of base excision repair enzyme activities glycosylase was examined in normal human skin fibroblasts (NHS) and to the induction of DNA replication and to cell division (9-11). fibroblasts from a patient with Bloom's syndrome. Using randomly In base excision repair, the initial enzymatic step is performed proliferating cells and those synchronized at specific intervals in the cell cycle, enzyme levels were shown to become elevated severalfold in a by a DNA glycosylase, which catalyzes the hydrolysis of the proliferation-associated manner. In NHS synchronized in ( .„byserum glycosyl bond releasing the modified base (2-4). Subsequent deprivation or in (., by isoleucine deprivation, maximal enzyme levels nuclease and DNA polymerase activities cleave and extend the were reached prior to maximal rates of DNA synthesis. In Bloom's resultant apurinic or apyrimidinic site in DNA and resynthesize syndrome cells synchronized in this manner, these two activities were the appropriate nucleotide sequences. Lastly, DNA ligase re coincident. Cells synchronized at the (.,-S border by hydroxyurea exhibit establishes the covalent linkage of the repaired DNA. an initial wave of DNA synthesis upon removal of the drug. The cells In normal human cells, uracil DNA glycosylase and the base then undergo another DNA synthetic cycle climaxing 18-21 h after excision repair pathway were enhanced prior to the induction release. Maximal hypoxanthine glycosylase activity of hydroxyurea- of DNA replication and DNA polymerase in serum-stimulated synchronized Bloom's cells was observed during the second round of cells (12, 13). We suggested that this enhancement of repair DNA synthesis. However, in NHS the peak of enzyme activity was enzyme activity might serve as a proofreading or prescreening observed as early as 9 h prior to the second round of DNA synthesis. To determine if hypoxanthine glycosylase could be induced in the absence mechanism to help prevent the fixation of mutagenic alterations of DNA synthesis, serum-synchronized NHS were released in the pres in the genome (14). Recently, we reported that hypermutable cells from Bloom's syndrome patients (15, 16) were character ence of hydroxyurea. The results showed that inhibition of DNA synthesis did not diminish glycosylase induction which demonstrated that DNA ized by an inability to correctly regulate the proliferation de replication was not required for glycosylase induction. pendent enhancement of base excision repair (17). In particular, uracil DNA glycosylase and the base excision repair pathway were enhanced coordinately with DNA replication. This would INTRODUCTION suggest that these individuals might be unable to efficiently repair carcinogen-modified DNA prior to replication. To ex Numerous studies have established the relationship between the initiation of neoplasia and the presence of chemical- or amine the specificity of this alteration in human gene expression radiation-induced DNA damage. DNA-carcinogen adducts we examined whether the regulation of another base excision such as 06-alkylguanine or <74-alkylthymidine are promutagenic repair enzyme, hypoxanthine DNA glycosylase, showed similar alterations in Bloom's syndrome cells. This enzyme excises and result in GC to AT transitions (1). Further, spontaneous deamination of cytidine to uracil and adenine to hypoxanthine hypoxanthine residues from DNA produced by the mutagenic represents mutational alterations in DNA. Enzymes which deamination of adenine (18). We now report that normal hu man cells enhanced this base excision repair enzyme prior to remove these lesions have been identified in mammalian cells DNA replication. However, in Bloom's syndrome cells, there (2-4). For the most part, their specific activities in diverse tissues correlate inversely with the susceptibility of such tissues was a temporal alteration such that the enzyme was induced to the action of carcinogenic agents. concomitantly with DNA replication. This altered temporal Eukaryotic cells specifically regulate the activities of diverse sequence was identical to that previously observed with the uracil DNA glycosylase. These results suggest that there is an DNA replicative enzymes during cell proliferation. In some intrinsic genetic defect in Bloom's syndrome cells which results instances, enzyme activities are induced to supply the increased quantities of metabolic intermediates for DNA synthesis. Thus, in the altered regulation of at least two individual base excision dihydrofolate reducíase, thymidine synthase, and thymidine repair enzymes during cell proliferation. kinase activities are increased as a function of growth (5-7). In contrast, in other instances, DNA replication and subsequent MATERIALS AND METHODS cell division would require the diminution of specific DNA Cell Culture. Normal human skin fibroblasts (CRL 1222; American metabolic enzymes; thus TTP dephosphorylase activity is di Type Culture Collection, Rockville, MD) and Bloom's syndrome fibro minished during cell proliferation (8). In each instance, the blasts (GM 2548; NIGMS Human Genetic Mutant Cell Repository, Camden, NJ) were obtained at passage 10 or lower. Cells were propa regulation of each gene would be an essential requirement for gated at 37°Cin 5% CO2 in air in 150-cm2 tissue culture flasks in DNA synthesis. Dulbecco's minimal essential medium (Gibco) containing glutamine, Similarly, the quantity of DNA repair enzymes in human antibiotics, and 10% fetal bovine serum unless specified. Cells were Received 10/25/85; revised 2/10/86, 4/1/86; accepted 4/16/86. subcultured after attaining confluence but were not maintained beyond The costs of publication of this article were defrayed in part by the payment passage IS. of page charges. This article must therefore be hereby marked advertisement in Methods of Cell Synchronization. Cells were synchronized in early accordance with 18 U.S.C. Section 1734 solely to indicate this fact. ' This study was supported by grants to M. A. S. from the W. W. Smith GI by serum, middle GI by isoleucine deprivation, or at the Gi-S border Charitable Trust, the National Science Foundation (DCB-8416295), and the NIH by incubation with 0.5 IHMHU.4 Cells to be synchronized by serum (CA-29414) and by grants to the Fels Research Institute from the NIH (CA 12227) and from the American Cancer Society (SIG-6). 4 The abbreviations used are: HU, hydroxyurea; NHS, normal human skin 2 Present address: Merrell Dow Research Institute, Cincinnati, OH 45215. fibroblasts; DMEM, Dulbecco's modified Eagle's medium; DTT, dithiothreitol; 3To whom requests for reprints should be addressed. HPLC, high-pressure liquid chromatography. 3756 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1986 American Association for Cancer Research. HYPOXANTHINE DNA GLYCOSYLASE REGULATION deprivation were seeded into 100 itiM plastic dishes at a density of IO6 cells per dish in DMEM containing 0.3% serum. After 5 days in culture, cells were released into the cell cycle by the addition of 20% serum containing medium. Synchronization by isoleucine deprivation was accomplished by seeding cells as above in 10% dialyzed fetal bovine serum containing DMEM without isoleucine (Gibco). After 3 days in culture, cells were released by the readdition of complete DMEM. HU synchronization of cultures was accomplished by the addition of a stock solution of HU to a final concentration of 0.5 HIM.After 18 h cells were rinsed twice with prewarmed Hanks' balanced salt solution (Gibco) and incubated with DMEM. For some experiments cells were given 2 miviHU immediately after release. For asynchronous growth, confluent cells in 150-cm2 tissue culture flasks were trypsinized and seeded in 100 mM dishes at a density of 1- 40 50 60 70 80 2 x 10s cells/dish. Cells were grown for 6 days without medium change. FRACTION NUMBER Cell Extracts. Cells to be used for various assays were rinsed twice Fig. 1. HPLC of ethanol-soluble products from hypoxanthine glycosylase with cold Dulbecco's phosphate-buffered saline (Gibco) and harvested assay. A typical chromatogram is shown. [3H]Deoxyinosine-containing substrate with a rubber policeman. After sedimentation at 300 x g, cell pellets was incubated with 10 ng of cell extract protein for 120 min at 37'C as described were frozen at -20°C.Prior to assay, frozen cells were resuspended in in "Materials and Methods." After precipitation with ethanol and carrier DNA, 1 ml of 20 mM Tris-HCl (pH 8.0)-1 mM DTT-20% (v/v) glycerol and the supernatant was concentrated under nitrogen. Authentic hypoxanthine was sonicated at 70 W for 30 s at 0°C,with a Bransonic sonicator equipped added, and the sample was injected directly on a Waters Ci8-f Bondapak reversed- phase column eluted isocratically at I ml/min with 10 mM ammonium formate, with a needle probe. Extracts were centrifuged at 300 x g to remove pH 3.8. Fractions were collected at 30-s intervals. The arrow indicates the position unbroken cells. of the hypoxanthine marker. Insel, release of hypoxanthine-specific radioactivity Preparation of Hypoxanthine DNA Glycosylase Substrate. Hypoxan- as a function of time of incubation (A. lit,.] of substrate and 10 ng of cell protein) or quantity of cell protein in assay mixture (O, 10 n\ of substrate, 10 »igofprotein, thine DNA glycosylase was assayed using a substrate prepared by a incubation for 120 min at 37"C). modi fil-aïionof a method reported by Karran (19). [3H]dATP (Amer- sham or ICN; specific activity, 26 Ci/mmol, 5.0 mCi) was deaminated say. Therefore, 1-2 Mgof hypoxanthine was used as a marker for 5 h with freshly prepared nitrous acid (3.0 Msodium nitrate adjusted to pH 3.9 with nitric acid).
Load more

Recommended publications
  • Human Hypoxanthine (Guanine) Phosphoribosyltransferase: An
    Proc. NatL Acad. Sci. USA Vol. 80, pp. 870-873, Febriary 1983 Medical Sciences Human hypoxanthine (guanine) phosphoribosyltransferase: An amino acid substitution in a mutant form of the enzyme isolated from a patient with gout (reverse-phase HPLC/peptide mapping/mutant enzyme) JAMES M. WILSON*t, GEORGE E. TARRt, AND WILLIAM N. KELLEY*t Departments of *Internal Medicine and tBiological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan 48109 Communicated by James B. Wyngaarden, November 3, 1982 ABSTRACT We have investigated the molecular basis for a tration ofenzyme protein. in both erythrocytes (3) and lympho- deficiency ofthe enzyme hypoxanthine (guanine) phosphoribosyl- blasts (4); (ii) a normal Vm., a normal Km for 5-phosphoribosyl- transferase (HPRT; IMP:pyrophosphate-phosphoribosyltransfer- 1-pyrophosphate, and a 5-fold increased Km for hypoxanthine ase, EC 2.4.2.8) in a patient with a severe form of gout. We re-. (unpublished data); (iii) a normal isoelectric point (3, 4) and ported in previous studies the isolation of a unique structural migration during nondenaturing polyacrylamide gel electro- variant of HPRT from this patient's erythrocytes and cultured phoresis (4); and (iv) -an apparently smaller subunit molecular lymphoblasts. This enzyme variant, which is called HPRTOnd0., weight as evidenced by an increased mobility during Na- is characterized by a decreased concentration of HPRT protein DodSO4/polyacrylamide gel electrophoresis (3, 4). in erythrocytes and lymphoblasts, a normal Vm.., a 5-fold in- Our study ofthe tryptic peptides and amino acid composition creased Km for hypoxanthine, a normal isoelectric point, and an of apparently smaller subunit molecular weight. Comparative pep- HPRTLondon revealed a single amino acid substitution (Ser tide mapping-experiments revealed a single abnormal tryptic pep- Leu) at position 109.
    [Show full text]
  • Studies on the Mechanism of Action of 6-Mercaptopurine in Sensitive and Resistant L1210 Leukemia in Vitro*
    Studies on the Mechanism of Action of 6-Mercaptopurine in Sensitive and Resistant L1210 Leukemia in Vitro* JACKD. DAVIDSON (Clinical Pharmacology and Experimental Tlierapeutics Service, National Cancer Institute, Betliesda, Md.) SUMMARY A study was made of the effects of 6-mercaptopurine (6-MP) upon nucleic acid metabolism in L1210 leukemia cells in vitro. Pharmacological concentrations of 6-MP inhibited the incorporation of both hypoxanthine and glycine into adenine nucleo- tides. Higher concentrations of 6-MP inhibited the incorporation of hypoxanthine into both adenine and guanine nucleotides. In a subline of L1210 which is resistant to 6-MP there was much less utilization of hypoxanthine than in the sensitive line, and incorporation into both adenine and guanine moieties was strongly inhibited by 6-MP. On the contrary, utilization of glycine for nucleotide purine synthesis was unaffected by 6-MP. These findings support the hypothesis that in L1210 leukemia 6-MP is metabolized to its ribotide, and this produces a metabolic block in the conversion of inosinic acid to adenylic acid. They further indicate that, in the L1210 cells resistant to 6-MP, there is a very limited capacity to convert 6-MP and hypoxanthine to ribotides. This leads to competition between 6-MP and hypoxanthine and to formation of insufficient 6-MP ribotide to cause the critical block. Although 6-mercaptopurine (6-MP) has been purines isolated from L1210 ascites leukemia cells studied with respect to its antitumor activity since have been studied. The results obtained suggest 1952, there is still relatively little information re that the primary antimetabolic activity of 6-MP garding the mechanism of its action.
    [Show full text]
  • A Previously Undescribed Pathway for Pyrimidine Catabolism
    A previously undescribed pathway for pyrimidine catabolism Kevin D. Loh*†, Prasad Gyaneshwar*‡, Eirene Markenscoff Papadimitriou*§, Rebecca Fong*, Kwang-Seo Kim*, Rebecca Parales¶, Zhongrui Zhouʈ, William Inwood*, and Sydney Kustu*,** *Department of Plant and Microbial Biology, 111 Koshland Hall, University of California, Berkeley, CA 94720-3102; ¶Section of Microbiology, 1 Shields Avenue, University of California, Davis, CA 95616; and ʈCollege of Chemistry, 8 Lewis Hall, University of California, Berkeley, CA 94720-1460 Contributed by Sydney Kustu, January 19, 2006 The b1012 operon of Escherichia coli K-12, which is composed of tive N sources. Here we present evidence that the b1012 operon seven unidentified ORFs, is one of the most highly expressed codes for proteins that constitute a previously undescribed operons under control of nitrogen regulatory protein C. Examina- pathway for pyrimidine degradation and thereby confirm the tion of strains with lesions in this operon on Biolog Phenotype view of Simaga and Kos (8, 9) that E. coli K-12 does not use either MicroArray (PM3) plates and subsequent growth tests indicated of the known pathways. that they failed to use uridine or uracil as the sole nitrogen source and that the parental strain could use them at room temperature Results but not at 37°C. A strain carrying an ntrB(Con) mutation, which Behavior on Biolog Phenotype MicroArray Plates. We tested our elevates transcription of genes under nitrogen regulatory protein parental strain NCM3722 and strains with mini Tn5 insertions in C control, could also grow on thymidine as the sole nitrogen several genes of the b1012 operon on Biolog (Hayward, CA) source, whereas strains with lesions in the b1012 operon could not.
    [Show full text]
  • Central Nervous System Dysfunction and Erythrocyte Guanosine Triphosphate Depletion in Purine Nucleoside Phosphorylase Deficiency
    Arch Dis Child: first published as 10.1136/adc.62.4.385 on 1 April 1987. Downloaded from Archives of Disease in Childhood, 1987, 62, 385-391 Central nervous system dysfunction and erythrocyte guanosine triphosphate depletion in purine nucleoside phosphorylase deficiency H A SIMMONDS, L D FAIRBANKS, G S MORRIS, G MORGAN, A R WATSON, P TIMMS, AND B SINGH Purine Laboratory, Guy's Hospital, London, Department of Immunology, Institute of Child Health, London, Department of Paediatrics, City Hospital, Nottingham, Department of Paediatrics and Chemical Pathology, National Guard King Khalid Hospital, Jeddah, Saudi Arabia SUMMARY Developmental retardation was a prominent clinical feature in six infants from three kindreds deficient in the enzyme purine nucleoside phosphorylase (PNP) and was present before development of T cell immunodeficiency. Guanosine triphosphate (GTP) depletion was noted in the erythrocytes of all surviving homozygotes and was of equivalent magnitude to that found in the Lesch-Nyhan syndrome (complete hypoxanthine-guanine phosphoribosyltransferase (HGPRT) deficiency). The similarity between the neurological complications in both disorders that the two major clinical consequences of complete PNP deficiency have differing indicates copyright. aetiologies: (1) neurological effects resulting from deficiency of the PNP enzyme products, which are the substrates for HGPRT, leading to functional deficiency of this enzyme. (2) immunodeficiency caused by accumulation of the PNP enzyme substrates, one of which, deoxyguanosine, is toxic to T cells. These studies show the need to consider PNP deficiency (suggested by the finding of hypouricaemia) in patients with neurological dysfunction, as well as in T cell immunodeficiency. http://adc.bmj.com/ They suggest an important role for GTP in normal central nervous system function.
    [Show full text]
  • Xanthine/Hypoxanthine Assay Kit
    Product Manual Xanthine/Hypoxanthine Assay Kit Catalog Number MET-5150 100 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Xanthine and hypoxanthine are naturally occurring purine derivatives. Xanthine is created from guanine by guanine deaminase, from hypoxanthine by xanthine oxidoreductase, and from xanthosine by purine nucleoside phosphorylase. Xanthine is used as a building block for human and animal drug medications, and is an ingredient in pesticides. In vitro, xanthine and related derivatives act as competitive nonselective phosphodiesterase inhibitors, raising intracellular cAMP, activating Protein Kinase A (PKA), inhibiting tumor necrosis factor alpha (TNF-α) as well as and leukotriene synthesis. Furthermore, xanthines can reduce levels of inflammation and act as nonselective adenosine receptor antagonists. Hypoxanthine is sometimes found in nucleic acids such as in the anticodon of tRNA in the form of its nucleoside inosine. Hypoxanthine is a necessary part of certain cell, bacteria, and parasitic cultures as a substrate and source of nitrogen. For example, hypoxanthine is often a necessary component in malaria parasite cultures, since Plasmodium falciparum needs hypoxanthine to make nucleic acids as well as to support energy metabolism. Recently NASA studies with meteorites found on Earth supported the idea that hypoxanthine and related organic molecules can be formed extraterrestrially. Hypoxanthine can form as a spontaneous deamination product of adenine. Because of its similar structure to guanine, the resulting hypoxanthine base can lead to an error in DNA transcription/replication, as it base pairs with cytosine. Hypoxanthine is typically removed from DNA by base excision repair and is initiated by N- methylpurine glycosylase (MPG).
    [Show full text]
  • Mechanism of Excessive Purine Biosynthesis in Hypoxanthine- Guanine Phosphoribosyltransferase Deficiency
    Mechanism of excessive purine biosynthesis in hypoxanthine- guanine phosphoribosyltransferase deficiency Leif B. Sorensen J Clin Invest. 1970;49(5):968-978. https://doi.org/10.1172/JCI106316. Research Article Certain gouty subjects with excessive de novo purine synthesis are deficient in hypoxanthineguanine phosphoribosyltransferase (HG-PRTase [EC 2.4.2.8]). The mechanism of accelerated uric acid formation in these patients was explored by measuring the incorporation of glycine-14C into various urinary purine bases of normal and enzyme-deficient subjects during treatment with the xanthine oxidase inhibitor, allopurinol. In the presence of normal HG-PRTase activity, allopurinol reduced purine biosynthesis as demonstrated by diminished excretion of total urinary purine or by reduction of glycine-14C incorporation into hypoxanthine, xanthine, and uric acid to less than one-half of control values. A boy with the Lesch-Nyhan syndrome was resistant to this effect of allopurinol while a patient with 12.5% of normal enzyme activity had an equivocal response. Three patients with normal HG-PRTase activity had a mean molar ratio of hypoxanthine to xanthine in the urine of 0.28, whereas two subjects who were deficient in HG-PRTase had reversal of this ratio (1.01 and 1.04). The patterns of 14C-labeling observed in HG-PRTase deficiency reflected the role of hypoxanthine as precursor of xanthine. The data indicate that excessive uric acid in HG-PRTase deficiency is derived from hypoxanthine which is insufficiently reutilized and, as a consequence thereof, catabolized inordinately to uric acid. The data provide evidence for cyclic interconversion of adenine and hypoxanthine derivatives. Cleavage of inosinic acid to hypoxanthine via inosine does […] Find the latest version: https://jci.me/106316/pdf Mechanism of Excessive Purine Biosynthesis in Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency LEIF B.
    [Show full text]
  • Non-Utilization of Radioactive Lodinated Uracil, Uridine, and Orotic Acid by Animal Tissues in Vivo W
    Non-utilization of Radioactive lodinated Uracil, Uridine, and Orotic Acid by Animal Tissues in Vivo W. H. PRUSOFF,WL. HOLMES,tANDA. D. WELCH Department of Pharmacology, School of Medicine, Western Reserve University, Cleveland, Ohio) Adenine (1, 6), guanine (1, 3, 5), cytidine (13), lometric localization of brain tumors. Further desoxycytidine (18), thymidine (18), and orotic more, an I'3-labeled oxazine dye had a significant acid (2), can be utilized by certain mammalian or effect in prolonging the life of mice bearing trans ganisms for the synthesis of nucleic acids ; and the planted tumors (19). If an effective and easily syn rate of incorporation of many of these compounds thesized radioactive iodine-labeled compound into the nucleic acids of rapidly growing tissues, could be found, the possibility might be afforded of such as regenerating liver or neoplastic tissues, is the comparable use of compounds labeled with greater than into those of resting tissues (10, 21). eka-iodine (astatine2@), a potent emitter of alpha Although 8-azaguanine is not a naturally occur particles, although this element is prepared with ring compound, evidence that it can be incorpo difficulty and has the inconveniently short half rated to a small extent into mammalian nucleic life of 7.5 hours (12). acids has been presented (16). This analog of gua Three P3-labeled pyrimidines, iodouridine-5- nine markedly inhibited the growth of Tetrahy I'S', iodouracil-5-I'31, and iodoorotic acid-5-P31, mena geleii, a guanine-requiring protozoan, and of were synthesized, and their incorporation into nor certain experimental tumors. Kidder et al.
    [Show full text]
  • Xj 128 IUMP Glucose Substance Will Be Provisionally Referred to As UDPX (Fig
    426 Studies on Uridine-Diphosphate-Glucose By A. C. PALADINI AND L. F. LELOIR Instituto de Inve8tigacione&s Bioquimicas, Fundacion Campomar, J. Alvarez 1719, Buenos Aires, Argentina (Received 18 September 1951) A previous paper (Caputto, Leloir, Cardini & found that the substance supposed to be uridine-2'- Paladini, 1950) reported the isolation of the co- phosphate was uridine-5'-phosphate. The hydrolysis enzyme of the galactose -1- phosphate --glucose - 1 - product of UDPG has now been compared with a phosphate transformation, and presented a tenta- synthetic specimen of uridine-5'-phosphate. Both tive structure for the substance. This paper deals substances were found to be identical as judged by with: (a) studies by paper chromatography of puri- chromatographic behaviour (Fig. 1) and by the rate fied preparations of uridine-diphosphate-glucose (UDPG); (b) the identification of uridine-5'-phos- 12A UDPG phate as a product of hydrolysis; (c) studies on the ~~~~~~~~~~~~~(a) alkaline degradation of UDPG, and (d) a substance similar to UDPG which will be referred to as UDPX. UMP Adenosine UDPG preparation8 8tudied by chromatography. 0 UjDPX Paper chromatography with appropriate solvents 0 has shown that some of the purest preparations of UDP UDPG which had been obtained previously contain two other compounds, uridinemonophosphate 0 4 (UMP) and a substance which appears to have the same constitution as UDPG except that it contains an unidentified component instead of glucose. This Xj 128 IUMP Glucose substance will be provisionally referred to as UDPX (Fig. la). The three components have been tested for co- enzymic activity in the galactose-1-phosphate-- 0-4 -J UDPX glucose-l-phosphate transformation, and it has been confirmed that UDPG is the active substance.
    [Show full text]
  • Availability and Quality of Published Data on the Purine Content of Foods, ⋆ T Alcoholic Beverages, and Dietary Supplements ⁎ Beiwen Wua, , Janet M
    Journal of Food Composition and Analysis 84 (2019) 103281 Contents lists available at ScienceDirect Journal of Food Composition and Analysis journal homepage: www.elsevier.com/locate/jfca Study Review Availability and quality of published data on the purine content of foods, ⋆ T alcoholic beverages, and dietary supplements ⁎ Beiwen Wua, , Janet M. Roselandb, David B. Haytowitzb,1, Pamela R. Pehrssonb, Abby G. Ershowc a Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA b Methods and Application of Food Composition Laboratory, Agricultural Research Service, US Department of Agriculture, Beltsville, MD 20705, USA c Office of Dietary Supplements, National Institutes of Health, Bethesda, MD 20892, USA ARTICLE INFO ABSTRACT Keywords: Gout, the most common type of inflammatory arthritis and associated with elevated uric acid levels, is aglobal Purine burden. “Western” dietary habits and lifestyle, and the resulting obesity epidemic, are often blamed for the Adenine increased prevalence of gout. Purine intake has shown the biggest dietary impact on uric acid. To manage this Guanine situation, data on the purine content of foods are needed. To assess availability and quality of purine data and Hypoxanthine identify research gaps, we obtained data for four purine bases (adenine, guanine, hypoxanthine, and xanthine) in Xanthine foods, alcoholic beverages, and dietary supplements. Data were predominantly from Japan, and very little from Gout Hyperuricemia the United States. Data quality was examined using a modified version of the USDA Data Quality Evaluation Food analysis System. Purine values in 298 foods/19 food groups, 15 alcoholic beverages, and 13 dietary supplements were Food composition reported. Mean hypoxanthine (mg/100 g) in the soups/sauces group was 112 and mean adenine in poultry Data quality organs was 62.4, which were the highest among all groups.
    [Show full text]
  • Developmental Disorder Associated with Increased Cellular Nucleotidase Activity (Purine-Pyrimidine Metabolism͞uridine͞brain Diseases)
    Proc. Natl. Acad. Sci. USA Vol. 94, pp. 11601–11606, October 1997 Medical Sciences Developmental disorder associated with increased cellular nucleotidase activity (purine-pyrimidine metabolismyuridineybrain diseases) THEODORE PAGE*†,ALICE YU‡,JOHN FONTANESI‡, AND WILLIAM L. NYHAN‡ Departments of *Neurosciences and ‡Pediatrics, University of California at San Diego, La Jolla, CA 92093 Communicated by J. Edwin Seegmiller, University of California at San Diego, La Jolla, CA, August 7, 1997 (received for review June 26, 1997) ABSTRACT Four unrelated patients are described with a represent defects of purine metabolism, although no specific syndrome that included developmental delay, seizures, ataxia, enzyme abnormality has been identified in these cases (6). In recurrent infections, severe language deficit, and an unusual none of these disorders has it been possible to delineate the behavioral phenotype characterized by hyperactivity, short mechanism through which the enzyme deficiency produces the attention span, and poor social interaction. These manifesta- neurological or behavioral abnormalities. Therapeutic strate- tions appeared within the first few years of life. Each patient gies designed to treat the behavioral and neurological abnor- displayed abnormalities on EEG. No unusual metabolites were malities of these disorders by replacing the supposed deficient found in plasma or urine, and metabolic testing was normal metabolites have not been successful in any case. except for persistent hypouricosuria. Investigation of purine This report describes four unrelated patients in whom and pyrimidine metabolism in cultured fibroblasts derived developmental delay, seizures, ataxia, recurrent infections, from these patients showed normal incorporation of purine speech deficit, and an unusual behavioral phenotype were bases into nucleotides but decreased incorporation of uridine.
    [Show full text]
  • Development, Uridine Diphosphate Glucose (UDPG), Pyrophosphorylase (EC 2.7.7.9)11 and Trehalose-6-Phosphate Synthetase (EC 2.3.1.15)
    PERIODS OF GENETIC TRANSCRIPTION REQUIRED FOR THE SYNTHESIS OF THREE ENZYMES DURING CELLULAR SLIME MOLD DEVELOPMENT* BY R. ROTH, t J. 1\I. ASHWORTH,4 AND M. SUSSMAN§ DEPARTMENT OF BIOLOGY, BRANDEIS UNIVERSITY, WALTHAM, MASSACHUSETTS Communicated by Sol Spiegelman, January 24, 1968 Various aspects of mRNA synthesis and stability have been examined in bacteria by permitting transcription to occur over a known, usually brief, period and then determining how much of a specific protein subsequently accumulates in the absence of further RNA synthesis. Thus, bacterial cells have been exposed to an inducer for very brief periods during which RNA synthesis could proceed normally and were then permitted to synthesize the enzyme de novo in the absence of both the inducer and further transcription. The latter restriction was ac- complished with base analogues,' uracil deprivation,2 actinomycin D,3 and by infection with lytic viruses.5 In an arginine- and uracil-requiring strain of E. coli infected with phage T6, protein and RNA synthesis were sequentially (and reversibly) restricted by successive precursor deprivations in order to study the accumulation of enzymes required for phage development.2 6 Under all of the above conditions, the amounts of enzymes synthesized were assumed to reflect in a simple fashion the over-all quantities and net concentrations of the correspond- irng mRNA species, and the subsequent decays of enzyme-forming capacity with time were assumed to reflect the manner in which the mRNA disappeared. This approach has also been exploited to investigate the transcriptive and translative events required for accumulation and disappearance of the enzyme uridine diphosphate galactose: polysaccharide transferase during slime mold development.
    [Show full text]
  • Nucleobases Thin Films Deposited on Nanostructured Transparent Conductive Electrodes for Optoelectronic Applications
    www.nature.com/scientificreports OPEN Nucleobases thin flms deposited on nanostructured transparent conductive electrodes for optoelectronic applications C. Breazu1*, M. Socol1, N. Preda1, O. Rasoga1, A. Costas1, G. Socol2, G. Petre1,3 & A. Stanculescu1* Environmentally-friendly bio-organic materials have become the centre of recent developments in organic electronics, while a suitable interfacial modifcation is a prerequisite for future applications. In the context of researches on low cost and biodegradable resource for optoelectronics applications, the infuence of a 2D nanostructured transparent conductive electrode on the morphological, structural, optical and electrical properties of nucleobases (adenine, guanine, cytosine, thymine and uracil) thin flms obtained by thermal evaporation was analysed. The 2D array of nanostructures has been developed in a polymeric layer on glass substrate using a high throughput and low cost technique, UV-Nanoimprint Lithography. The indium tin oxide electrode was grown on both nanostructured and fat substrate and the properties of the heterostructures built on these two types of electrodes were analysed by comparison. We report that the organic-electrode interface modifcation by nano- patterning afects both the optical (transmission and emission) properties by multiple refections on the walls of nanostructures and the electrical properties by the efect on the organic/electrode contact area and charge carrier pathway through electrodes. These results encourage the potential application of the nucleobases thin flms deposited on nanostructured conductive electrode in green optoelectronic devices. Te use of natural or nature-inspired materials in organic electronics is a dynamic emerging research feld which aims to replace the synthesized materials with natural (bio) ones in organic electronics1–3.
    [Show full text]