guanosine-triphosphate and Anemia

Genetic variation of inosine triphosphatase (ITPA) causing an accumulation of inosine triphosphate (ITP) has been shown to protect patients against ribavirin (RBV)-induced anemia during treatment for chronic hepatitis C infection by genome-wide association study (GWAS). However, the biologic mechanism by which this occurs is unknown.. We examined whether ITP can be used by adenosine triphosphatase (ATPase) in human erythrocytes or recombinant human adenylosuccinate synthase (ADSS). RBV-induced adenosine triphosphate (ATP) reduction in erythrocytes was compared with the genetically determined low or normal activity of ITPA, leading respectively to high or normal ITP levels.. Although ITP is not used directly by human erythrocyte ATPase, it can be used for ATP biosynthesis via ADSS in place of guanosine triphosphate (GTP). With RBV challenge, erythrocyte ATP reduction was more severe in the wild-type ITPA genotype than in the hemolysis protective ITPA genotype. This difference also remains after inhibiting adenosine uptake using nitrobenzylmercaptopurine riboside (NBMPR). Interestingly, the alleviation of ATP reduction by the hemolysis protective ITPA genotype was canceled by the ADSS inhibitor 6-mercaptoethanol (6-MP).. ITP confers protection against RBV-induced ATP reduction by substituting for erythrocyte GTP, which is depleted by RBV, in the biosynthesis of ATP. Because patients with excess ITP appear largely protected against anemia, these results confirm that RBV-induced anemia is due primarily to the effect of the drug on GTP and consequently ATP levels in erythrocytes.

Topics: Adenosine Triphosphate; Adenylosuccinate Synthase; Adolescent; Adult; Anemia; Antiviral Agents; Enzyme Activation; Erythrocytes; Genetic Variation; Genotype; Guanosine Triphosphate; Hepatitis C, Chronic; Humans; In Vitro Techniques; Inosine Triphosphatase; Inosine Triphosphate; Pyrophosphatases; Ribavirin; Young Adult

To explore the mechanism of erythropoietin action on differentiation of erythroblasts, we have examined its effect on regulating phosphorylation of the 25-kD mRNA cap binding protein (eIF-4E). Erythroblasts from the spleens of mice infected with the anemia strain of Friend virus (FVA cells) were studied. Erythropoietin stimulated phosphorylation of eIF-4E in FVA cells within 30 minutes, and this effect was maximal at 60 minutes. Phosphoamino acid analysis and tryptic phosphopeptide map analysis of eIF-4E isolated from both control and erythropoietin-treated cells identified a predominant phosphopeptide containing phosphoserine. However, when cells were incubated with 1 muM okadaic acid, eIF-4E was phosphorylated on both serine and threonine residues and three additional tryptic phosphopeptides were detected. We also identified a 37-kD phosphoprotein (pp37) that bound specifically to the m7GTP cap structure and coimmunoprecipitated with eIF-kD protein was phosphorylated on both serine and threonine residues. These results indicate that phosphorylation of eIF-4E is a target in erythropoietin-initiated signal transduction events and that this phosphorylation precedes observable effects of erythropoietin on macromolecular biosynthesis. Although of pp37 remains to be studied, it may represent a developmentally regulated mRNA cap binding protein.

Topics: Anemia; Animals; Erythroblasts; Erythropoietin; Ethers, Cyclic; Eukaryotic Initiation Factor-4E; Female; Friend murine leukemia virus; Guanosine Diphosphate; Guanosine Triphosphate; Mice; Molecular Weight; Okadaic Acid; Peptide Initiation Factors; Phosphoproteins; Phosphorylation; RNA Caps; RNA, Messenger; Spleen; Tetradecanoylphorbol Acetate

The effects of two different degrees of experimentally induced anemia and the consequent high percentage of circulating immature erythrocytes on oxygen affinity (pH 7.5 and 41 degrees C), erythrocyte organic phosphates, and Hb fractions have been studied in quail. Blood reticulocytes reached percentages of 24 and 69-87% in the first and second experiments, respectively. Variations in the phosphate levels found during the anemic process were related to the amount of circulating reticulocytes. The erythrocyte [ATP] (brackets indicate concentration) and [ATP]/[Hb] molar ratio increased with the release of reticulocytes and returned to control levels as they matured. The erythrocyte [inositol pentakisphosphate (InsP5)] decreased significantly when circulating reticulocytes showed their highest values, whereas there was hardly any effect on the [InsP5]/[Hb] molar ratio, which changed only slightly. Hb-O2 affinity also exhibited no statistical changes associated with acute anemia. These latter findings indicate that InsP5, at physiological concentrations, is the primary modulator of quail Hb function; the observed rise in [ATP] has no additional influence on Hb-O2 affinity. It is suggested that InsP5 tends to maintain the blood oxygen affinity in both mature erythrocytes and reticulocytes. The main compensatory response at blood level is a rapid bulk reticulocyte release from medulla.

Topics: Adenosine Triphosphate; Anemia; Animals; Biological Transport; Coturnix; Erythrocyte Count; Erythrocyte Indices; Erythrocytes; Guanosine Triphosphate; Hematocrit; Hemoglobins; Oxygen; Phenylhydrazines; Phosphates; Reticulocytes

Topics: Adenosine Triphosphate; Anemia; Animals; Cattle; Cytidine Triphosphate; Erythrocytes; Guanosine Triphosphate; Kinetics; Male; Phenylhydrazines; Reticulocytes; Ribonucleosides; Ribonucleotides; Uric Acid; Uridine Triphosphate