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. 2000 Apr 1;347(Pt 1):233–242.

Cytokine-inducible enhancer with promoter activity in both the rat and human manganese-superoxide dismutase genes.

R J Rogers 1, S E Chesrown 1, S Kuo 1, J M Monnier 1, H S Nick 1
PMCID: PMC1220953  PMID: 10727424

Abstract

Diverse pro-inflammatory mediators regulate transcription of the gene (MnSOD) encoding the mitochondrial anti-oxidant protein manganese-superoxide dismutase. Understanding the regulation of this gene is crucial to comprehending its role in cytoprotection. In transfected lung epithelial cells, a human-growth-hormone reporter gene system was utilized to identify a potential enhancer in the MnSOD genomic fragment previously shown to contain multiple DNase-I-hypersensitive sites. Northern analysis demonstrated a 10-20-fold increase in response to pro-inflammatory mediators. Inclusion of the MnSOD genomic fragment in reporter constructs was necessary to mimic these stimulus-dependent endogenous levels. The inducible enhancer element was localized to a 260 bp fragment in intron 2, coinciding with a previously defined DNase-I-hypersensitive site. This element functions in an orientation- and position-independent manner as well as with the heterologous thymidine kinase promoter. In addition, we have demonstrated that a homologous sequence within the human MnSOD gene exhibits identical enhancer activity. A novel characteristic of the rat and human enhancer elements involves the ability to promote cytokine-inducible transcription in the absence of a classical promoter.

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Selected References

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  1. Akashi M., Hachiya M., Paquette R. L., Osawa Y., Shimizu S., Suzuki G. Irradiation increases manganese superoxide dismutase mRNA levels in human fibroblasts. Possible mechanisms for its accumulation. J Biol Chem. 1995 Jun 30;270(26):15864–15869. doi: 10.1074/jbc.270.26.15864. [DOI] [PubMed] [Google Scholar]
  2. Andrews N. C., Faller D. V. A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res. 1991 May 11;19(9):2499–2499. doi: 10.1093/nar/19.9.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker K., Marcus C. B., Huffman K., Kruk H., Malfroy B., Doctrow S. R. Synthetic combined superoxide dismutase/catalase mimetics are protective as a delayed treatment in a rat stroke model: a key role for reactive oxygen species in ischemic brain injury. J Pharmacol Exp Ther. 1998 Jan;284(1):215–221. [PubMed] [Google Scholar]
  4. Callis J., Fromm M., Walbot V. Introns increase gene expression in cultured maize cells. Genes Dev. 1987 Dec;1(10):1183–1200. doi: 10.1101/gad.1.10.1183. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Del Maestro R., McDonald W. Subcellular localization of superoxide dismutases, glutathione peroxidase and catalase in developing rat cerebral cortex. Mech Ageing Dev. 1989 Apr;48(1):15–31. doi: 10.1016/0047-6374(89)90022-5. [DOI] [PubMed] [Google Scholar]
  7. Dougall W. C., Nick H. S. Manganese superoxide dismutase: a hepatic acute phase protein regulated by interleukin-6 and glucocorticoids. Endocrinology. 1991 Nov;129(5):2376–2384. doi: 10.1210/endo-129-5-2376. [DOI] [PubMed] [Google Scholar]
  8. Dougall W. C., Nick H. S. Manganese superoxide dismutase: a hepatic acute phase protein regulated by interleukin-6 and glucocorticoids. Endocrinology. 1991 Nov;129(5):2376–2384. doi: 10.1210/endo-129-5-2376. [DOI] [PubMed] [Google Scholar]
  9. Eastgate J., Moreb J., Nick H. S., Suzuki K., Taniguchi N., Zucali J. R. A role for manganese superoxide dismutase in radioprotection of hematopoietic stem cells by interleukin-1. Blood. 1993 Feb 1;81(3):639–646. [PubMed] [Google Scholar]
  10. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fujii J., Taniguchi N. Phorbol ester induces manganese-superoxide dismutase in tumor necrosis factor-resistant cells. J Biol Chem. 1991 Dec 5;266(34):23142–23146. [PubMed] [Google Scholar]
  12. Halliwell B., Gutteridge J. M. Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol. 1990;186:1–85. doi: 10.1016/0076-6879(90)86093-b. [DOI] [PubMed] [Google Scholar]
  13. Harris C. A., Derbin K. S., Hunte-McDonough B., Krauss M. R., Chen K. T., Smith D. M., Epstein L. B. Manganese superoxide dismutase is induced by IFN-gamma in multiple cell types. Synergistic induction by IFN-gamma and tumor necrosis factor or IL-1. J Immunol. 1991 Jul 1;147(1):149–154. [PubMed] [Google Scholar]
  14. Hurt J., Hsu J. L., Dougall W. C., Visner G. A., Burr I. M., Nick H. S. Multiple mRNA species generated by alternate polyadenylation from the rat manganese superoxide dismutase gene. Nucleic Acids Res. 1992 Jun 25;20(12):2985–2990. doi: 10.1093/nar/20.12.2985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones P. L., Ping D., Boss J. M. Tumor necrosis factor alpha and interleukin-1beta regulate the murine manganese superoxide dismutase gene through a complex intronic enhancer involving C/EBP-beta and NF-kappaB. Mol Cell Biol. 1997 Dec;17(12):6970–6981. doi: 10.1128/mcb.17.12.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Judde J. G., Max E. E. Characterization of the human immunoglobulin kappa gene 3' enhancer: functional importance of three motifs that demonstrate B-cell-specific in vivo footprints. Mol Cell Biol. 1992 Nov;12(11):5206–5216. doi: 10.1128/mcb.12.11.5206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuo S., Chesrown S. E., Mellott J. K., Rogers R. J., Hsu J. L., Nick H. S. In vivo architecture of the manganese superoxide dismutase promoter. J Biol Chem. 1999 Feb 5;274(6):3345–3354. doi: 10.1074/jbc.274.6.3345. [DOI] [PubMed] [Google Scholar]
  18. Li Y., Huang T. T., Carlson E. J., Melov S., Ursell P. C., Olson J. L., Noble L. J., Yoshimura M. P., Berger C., Chan P. H. Dilated cardiomyopathy and neonatal lethality in mutant mice lacking manganese superoxide dismutase. Nat Genet. 1995 Dec;11(4):376–381. doi: 10.1038/ng1295-376. [DOI] [PubMed] [Google Scholar]
  19. Manganaro F., Chopra V. S., Mydlarski M. B., Bernatchez G., Schipper H. M. Redox perturbations in cysteamine-stressed astroglia: implications for inclusion formation and gliosis in the aging brain. Free Radic Biol Med. 1995 Dec;19(6):823–835. doi: 10.1016/0891-5849(95)02008-x. [DOI] [PubMed] [Google Scholar]
  20. Melov S., Schneider J. A., Day B. J., Hinerfeld D., Coskun P., Mirra S. S., Crapo J. D., Wallace D. C. A novel neurological phenotype in mice lacking mitochondrial manganese superoxide dismutase. Nat Genet. 1998 Feb;18(2):159–163. doi: 10.1038/ng0298-159. [DOI] [PubMed] [Google Scholar]
  21. Ptashne M. Gene regulation by proteins acting nearby and at a distance. Nature. 1986 Aug 21;322(6081):697–701. doi: 10.1038/322697a0. [DOI] [PubMed] [Google Scholar]
  22. Queen C., Baltimore D. Immunoglobulin gene transcription is activated by downstream sequence elements. Cell. 1983 Jul;33(3):741–748. doi: 10.1016/0092-8674(83)90016-8. [DOI] [PubMed] [Google Scholar]
  23. Selden R. F., Howie K. B., Rowe M. E., Goodman H. M., Moore D. D. Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol Cell Biol. 1986 Sep;6(9):3173–3179. doi: 10.1128/mcb.6.9.3173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shields J. M., Yang V. W. Identification of the DNA sequence that interacts with the gut-enriched Krüppel-like factor. Nucleic Acids Res. 1998 Feb 1;26(3):796–802. doi: 10.1093/nar/26.3.796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shull S., Heintz N. H., Periasamy M., Manohar M., Janssen Y. M., Marsh J. P., Mossman B. T. Differential regulation of antioxidant enzymes in response to oxidants. J Biol Chem. 1991 Dec 25;266(36):24398–24403. [PubMed] [Google Scholar]
  26. Simkevich C. P., Thompson J. P., Poppleton H., Raghow R. The transcriptional tissue specificity of the human pro alpha 1 (I) collagen gene is determined by a negative cis-regulatory element in the promoter. Biochem J. 1992 Aug 15;286(Pt 1):179–185. doi: 10.1042/bj2860179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Suzuki K., Tatsumi H., Satoh S., Senda T., Nakata T., Fujii J., Taniguchi N. Manganese-superoxide dismutase in endothelial cells: localization and mechanism of induction. Am J Physiol. 1993 Oct;265(4 Pt 2):H1173–H1178. doi: 10.1152/ajpheart.1993.265.4.H1173. [DOI] [PubMed] [Google Scholar]
  28. Takimoto Y., Kuramoto A. Presence of a regulatory element within the first intron of the human platelet-derived growth factor-A chain gene. Jpn J Cancer Res. 1993 Dec;84(12):1268–1272. doi: 10.1111/j.1349-7006.1993.tb02833.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Valentine J. F., Nick H. S. Acute-phase induction of manganese superoxide dismutase in intestinal epithelial cell lines. Gastroenterology. 1992 Sep;103(3):905–912. doi: 10.1016/0016-5085(92)90024-s. [DOI] [PubMed] [Google Scholar]
  30. Visner G. A., Block E. R., Burr I. M., Nick H. S. Regulation of manganese superoxide dismutase in porcine pulmonary artery endothelial cells. Am J Physiol. 1991 Jun;260(6 Pt 1):L444–L449. doi: 10.1152/ajplung.1991.260.6.L444. [DOI] [PubMed] [Google Scholar]
  31. Visner G. A., Chesrown S. E., Monnier J., Ryan U. S., Nick H. S. Regulation of manganese superoxide dismutase: IL-1 and TNF induction in pulmonary artery and microvascular endothelial cells. Biochem Biophys Res Commun. 1992 Oct 15;188(1):453–462. doi: 10.1016/0006-291x(92)92406-n. [DOI] [PubMed] [Google Scholar]
  32. Visner G. A., Dougall W. C., Wilson J. M., Burr I. A., Nick H. S. Regulation of manganese superoxide dismutase by lipopolysaccharide, interleukin-1, and tumor necrosis factor. Role in the acute inflammatory response. J Biol Chem. 1990 Feb 15;265(5):2856–2864. [PubMed] [Google Scholar]
  33. Wispé J. R., Warner B. B., Clark J. C., Dey C. R., Neuman J., Glasser S. W., Crapo J. D., Chang L. Y., Whitsett J. A. Human Mn-superoxide dismutase in pulmonary epithelial cells of transgenic mice confers protection from oxygen injury. J Biol Chem. 1992 Nov 25;267(33):23937–23941. [PubMed] [Google Scholar]
  34. Wong G. H., Elwell J. H., Oberley L. W., Goeddel D. V. Manganous superoxide dismutase is essential for cellular resistance to cytotoxicity of tumor necrosis factor. Cell. 1989 Sep 8;58(5):923–931. doi: 10.1016/0092-8674(89)90944-6. [DOI] [PubMed] [Google Scholar]
  35. Wong G. H., Goeddel D. V. Induction of manganous superoxide dismutase by tumor necrosis factor: possible protective mechanism. Science. 1988 Nov 11;242(4880):941–944. doi: 10.1126/science.3263703. [DOI] [PubMed] [Google Scholar]
  36. Zhang W., Shields J. M., Sogawa K., Fujii-Kuriyama Y., Yang V. W. The gut-enriched Krüppel-like factor suppresses the activity of the CYP1A1 promoter in an Sp1-dependent fashion. J Biol Chem. 1998 Jul 10;273(28):17917–17925. doi: 10.1074/jbc.273.28.17917. [DOI] [PMC free article] [PubMed] [Google Scholar]

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