Hypoxia regulates xanthine dehydrogenase activity at pre- and posttranslational levels.

Hypoxia increases the activity of xanthine oxidase (XO) and its precursor, xanthine dehydrogenase (XDH), but the mechanism of regulation is unclear. In hypoxic Swiss 3T3 cells, an early (0-24 h) cycloheximide-insensitive increase in XO-XDH activity, coupled with a lack of increase in de novo XO-XDH synthesis (immunoprecipitation) or mRNA levels (quantitative RT-PCR), demonstrated a posttranslational effect of hypoxia. Similarly, hyperoxia decreased XO-XDH activity faster than could be accounted for by cessation of XO-XDH protein synthesis. In further support of a posttranslational effect, cells transfected with a constitutively driven XDH construct displayed an exaggerated increase in activity in hypoxia but no increase in activity in hyperoxia. However, more prolonged exposure to hypoxia (24-48 h) induced an increase in XO-XDH mRNA levels and de novo XO-XDH protein synthesis, suggesting an additional pretranslational effect. Finally, hypoxic induction of XO-XDH activity was found to be cell-type-restricted. We conclude that control of XO-XDH levels by oxygen tension is a complex process which involves several points of regulation.

[1]  P. Hassoun,et al.  Determination of xanthine dehydrogenase mRNA by a reverse transcription-coupled competitive quantitative polymerase chain reaction assay: regulation in rat endothelial cells by hypoxia and hyperoxia. , 1996, Archives of biochemistry and biophysics.

[2]  J. Hoidal,et al.  Molecular cloning and characterization of the human xanthine dehydrogenase gene (XDH). , 1996, Genomics.

[3]  K. Raivio,et al.  Cloning and expression in vitro of human xanthine dehydrogenase/oxidase. , 1996, The Biochemical journal.

[4]  V. Shepherd,et al.  Nitric oxide inactivates xanthine dehydrogenase and xanthine oxidase in interferon-gamma-stimulated macrophages. , 1994, American journal of respiratory cell and molecular biology.

[5]  G. Semenza,et al.  Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1. , 1994, The Journal of biological chemistry.

[6]  M. Seldin,et al.  Chromosomal mapping, isolation, and characterization of the mouse xanthine dehydrogenase gene. , 1994, Genomics.

[7]  K. Pfeffer,et al.  Xanthine dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells. Cytokine and steroid regulation. , 1994, Journal of immunology.

[8]  S. Ozawa,et al.  Thyroxine increases the levels of epidermal growth factor messenger ribonucleic acid (EGF mRNA) in the thyroid in vivo, as revealed by quantitative reverse transcription polymerase chain reaction with an internal control EGF mRNA. , 1993, Endocrinology.

[9]  R. Harrison,et al.  Purification of xanthine oxidase from human heart. , 1993, Biochemical Society transactions.

[10]  P. Ratcliffe,et al.  Inducible operation of the erythropoietin 3' enhancer in multiple cell lines: evidence for a widespread oxygen-sensing mechanism. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[11]  P. Casellas,et al.  Standardization of mRNA titration using a polymerase chain reaction method involving co-amplification with a multispecific internal control. , 1992, The Journal of biological chemistry.

[12]  F. Falciani,et al.  Interferons induce xanthine dehydrogenase gene expression in L929 cells. , 1992, The Biochemical journal.

[13]  M. Benboubetra,et al.  Purification and partial characterization of xanthine oxidase from human milk. , 1992, Biochimica et biophysica acta.

[14]  B. Freeman,et al.  The contribution of vascular endothelial xanthine dehydrogenase/oxidase to oxygen-mediated cell injury. , 1992, Archives of biochemistry and biophysics.

[15]  P. Perani,et al.  Molecular cloning of a cDNA coding for mouse liver xanthine dehydrogenase. Regulation of its transcript by interferons in vivo. , 1992, The Biochemical journal.

[16]  J. Repine,et al.  Hypoxia injures endothelial cells by increasing endogenous xanthine oxidase activity. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[17]  U. Ryan,et al.  Regulation of xanthine dehydrogenase and xanthine oxidase activity and gene expression in cultured rat pulmonary endothelial cells. , 1992, The Journal of clinical investigation.

[18]  J. Repine,et al.  Inactivation of xanthine oxidase by hydrogen peroxide involves site-directed hydroxyl radical formation. , 1991, Free radical biology & medicine.

[19]  M. Sato,et al.  Proteolytic conversion of xanthine dehydrogenase from the NAD-dependent type to the O2-dependent type. Amino acid sequence of rat liver xanthine dehydrogenase and identification of the cleavage sites of the enzyme protein during irreversible conversion by trypsin. , 1990, The Journal of biological chemistry.

[20]  A. Harken,et al.  Hyperoxia and self- or neutrophil-generated O2 metabolites inactivate xanthine oxidase. , 1988, Journal of applied physiology.

[21]  H. P. Jones,et al.  Conversion of xanthine dehydrogenase to oxidase in ischemic rat tissues. , 1987, The Journal of clinical investigation.

[22]  W. Bender,et al.  Mutations affecting expression of the rosy locus in Drosophila melanogaster. , 1987, Genetics.

[23]  R. Lewontin,et al.  Sequence of the structural gene for xanthine dehydrogenase (rosy locus) in Drosophila melanogaster. , 1987, Genetics.

[24]  J. McCord,et al.  Oxygen-derived free radicals in postischemic tissue injury. , 1985, The New England journal of medicine.

[25]  K. Rajagopalan,et al.  Drosophila melanogaster ma-l mutants are defective in the sulfuration of desulfo Mo hydroxylases. , 1982, The Journal of biological chemistry.

[26]  E. Jaffe,et al.  Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. , 1973, The Journal of clinical investigation.

[27]  G. Kikuchi,et al.  Differential induction of fraction I and fraction II of delta-aminolevulinate synthetase in Rhodopseudomonas spheroides under various incubation conditions. , 1970, Archives of biochemistry and biophysics.