Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Mar 15;90(6):2423–2427. doi: 10.1073/pnas.90.6.2423

Inducible operation of the erythropoietin 3' enhancer in multiple cell lines: evidence for a widespread oxygen-sensing mechanism.

P H Maxwell 1, C W Pugh 1, P J Ratcliffe 1
PMCID: PMC46099  PMID: 8460154

Abstract

Adaptive responses to hypoxia occur in many biological systems. A well-characterized example is the hypoxic induction of the synthesis of erythropoietin, a hormone which regulates erythropoiesis and hence blood oxygen content. The restricted expression of the erythropoietin gene in subsets of cells within kidney and liver has suggested that this specific oxygen-sensing mechanism is restricted to specialized cells in those organs. Using transient transfection of reporter genes coupled to a transcriptional enhancer lying 3' to the erythropoietin gene, we show that an oxygen-sensing system similar, or identical, to that controlling erythropoietin expression is wide-spread in mammalian cells. The extensive distribution of this sensing mechanism contrasts with the restricted expression of erythropoietin, suggesting that it mediates other adaptive responses to hypoxia.

Full text

PDF

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ager A. Isolation and culture of high endothelial cells from rat lymph nodes. J Cell Sci. 1987 Feb;87(Pt 1):133–144. doi: 10.1242/jcs.87.1.133. [DOI] [PubMed] [Google Scholar]
  2. Beck I., Ramirez S., Weinmann R., Caro J. Enhancer element at the 3'-flanking region controls transcriptional response to hypoxia in the human erythropoietin gene. J Biol Chem. 1991 Aug 25;266(24):15563–15566. [PubMed] [Google Scholar]
  3. Benjamin I. J., Kröger B., Williams R. S. Activation of the heat shock transcription factor by hypoxia in mammalian cells. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6263–6267. doi: 10.1073/pnas.87.16.6263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beru N., McDonald J., Lacombe C., Goldwasser E. Expression of the erythropoietin gene. Mol Cell Biol. 1986 Jul;6(7):2571–2575. doi: 10.1128/mcb.6.7.2571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Conkie D., Affara N., Harrison P. R., Paul J., Jones K. In situ localization of globin messenger RNA formation. II. After treatment of Friend virus-transformed mouse cells with dimethyl sulfoxide. J Cell Biol. 1974 Nov;63(2 Pt 1):414–419. doi: 10.1083/jcb.63.2.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goldberg M. A., Dunning S. P., Bunn H. F. Regulation of the erythropoietin gene: evidence that the oxygen sensor is a heme protein. Science. 1988 Dec 9;242(4884):1412–1415. doi: 10.1126/science.2849206. [DOI] [PubMed] [Google Scholar]
  7. Goldberg M. A., Glass G. A., Cunningham J. M., Bunn H. F. The regulated expression of erythropoietin by two human hepatoma cell lines. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7972–7976. doi: 10.1073/pnas.84.22.7972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imagawa S., Goldberg M. A., Doweiko J., Bunn H. F. Regulatory elements of the erythropoietin gene. Blood. 1991 Jan 15;77(2):278–285. [PubMed] [Google Scholar]
  9. Knighton D. R., Hunt T. K., Scheuenstuhl H., Halliday B. J., Werb Z., Banda M. J. Oxygen tension regulates the expression of angiogenesis factor by macrophages. Science. 1983 Sep 23;221(4617):1283–1285. doi: 10.1126/science.6612342. [DOI] [PubMed] [Google Scholar]
  10. Kourembanas S., Marsden P. A., McQuillan L. P., Faller D. V. Hypoxia induces endothelin gene expression and secretion in cultured human endothelium. J Clin Invest. 1991 Sep;88(3):1054–1057. doi: 10.1172/JCI115367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Koury S. T., Bondurant M. C., Koury M. J. Localization of erythropoietin synthesizing cells in murine kidneys by in situ hybridization. Blood. 1988 Feb;71(2):524–527. [PubMed] [Google Scholar]
  12. Koury S. T., Bondurant M. C., Koury M. J., Semenza G. L. Localization of cells producing erythropoietin in murine liver by in situ hybridization. Blood. 1991 Jun 1;77(11):2497–2503. [PubMed] [Google Scholar]
  13. Krantz S. B. Erythropoietin. Blood. 1991 Feb 1;77(3):419–434. [PubMed] [Google Scholar]
  14. Lacombe C., Da Silva J. L., Bruneval P., Fournier J. G., Wendling F., Casadevall N., Camilleri J. P., Bariety J., Varet B., Tambourin P. Peritubular cells are the site of erythropoietin synthesis in the murine hypoxic kidney. J Clin Invest. 1988 Feb;81(2):620–623. doi: 10.1172/JCI113363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Necas E., Thorling E. B. Unresponsiveness of erythropoietin-producing cells to cyanide. Am J Physiol. 1972 May;222(5):1187–1190. doi: 10.1152/ajplegacy.1972.222.5.1187. [DOI] [PubMed] [Google Scholar]
  16. Pugh C. W., Tan C. C., Jones R. W., Ratcliffe P. J. Functional analysis of an oxygen-regulated transcriptional enhancer lying 3' to the mouse erythropoietin gene. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10553–10557. doi: 10.1073/pnas.88.23.10553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  18. Semenza G. L., Koury S. T., Nejfelt M. K., Gearhart J. D., Antonarakis S. E. Cell-type-specific and hypoxia-inducible expression of the human erythropoietin gene in transgenic mice. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8725–8729. doi: 10.1073/pnas.88.19.8725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Semenza G. L., Nejfelt M. K., Chi S. M., Antonarakis S. E. Hypoxia-inducible nuclear factors bind to an enhancer element located 3' to the human erythropoietin gene. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5680–5684. doi: 10.1073/pnas.88.13.5680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Semenza G. L., Traystman M. D., Gearhart J. D., Antonarakis S. E. Polycythemia in transgenic mice expressing the human erythropoietin gene. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2301–2305. doi: 10.1073/pnas.86.7.2301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shoemaker C. B., Mitsock L. D. Murine erythropoietin gene: cloning, expression, and human gene homology. Mol Cell Biol. 1986 Mar;6(3):849–858. doi: 10.1128/mcb.6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tan C. C., Eckardt K. U., Firth J. D., Ratcliffe P. J. Feedback modulation of renal and hepatic erythropoietin mRNA in response to graded anemia and hypoxia. Am J Physiol. 1992 Sep;263(3 Pt 2):F474–F481. doi: 10.1152/ajprenal.1992.263.3.F474. [DOI] [PubMed] [Google Scholar]
  23. Tan C. C., Ratcliffe P. J. Effect of inhibitors of oxidative phosphorylation on erythropoietin mRNA in isolated perfused rat kidneys. Am J Physiol. 1991 Dec;261(6 Pt 2):F982–F987. doi: 10.1152/ajprenal.1991.261.6.F982. [DOI] [PubMed] [Google Scholar]
  24. Webster K. A., Gunning P., Hardeman E., Wallace D. C., Kedes L. Coordinate reciprocal trends in glycolytic and mitochondrial transcript accumulations during the in vitro differentiation of human myoblasts. J Cell Physiol. 1990 Mar;142(3):566–573. doi: 10.1002/jcp.1041420316. [DOI] [PubMed] [Google Scholar]
  25. Westerveld A., Visser R. P., Meera Khan P., Bootsma D. Loss of human genetic markers in man--Chinese hamster somatic cell hybrids. Nat New Biol. 1971 Nov 3;234(44):20–24. doi: 10.1038/newbio234020a0. [DOI] [PubMed] [Google Scholar]
  26. Widelitz R. B., Magun B. E., Gerner E. W. Effects of cycloheximide on thermotolerance expression, heat shock protein synthesis, and heat shock protein mRNA accumulation in rat fibroblasts. Mol Cell Biol. 1986 Apr;6(4):1088–1094. doi: 10.1128/mcb.6.4.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zitomer R. S., Lowry C. V. Regulation of gene expression by oxygen in Saccharomyces cerevisiae. Microbiol Rev. 1992 Mar;56(1):1–11. doi: 10.1128/mr.56.1.1-11.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES