Skip to main content

Advertisement

Log in

Ets-1 is involved in transcriptional regulation of the chick inducible nitric oxide synthase gene in embryonic ventricular myoctyes

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

In order to elucidate roles of Ets family of transcription factors in transcriptional activation of inducible nitric oxide synthase (iNOS) genes, we analyzed the chick iNOS gene expression in cultured chick embryonic ventricular myocytes (CEVM). Deletional analysis and site-directed mutagenesis demonstrated that both the Ets/PEA3 site (–221 to –216 bp) and the κB site (–101 to –93 bp) of the 5′-flanking region of the chick iNOS gene were involved in the maximal activation of the lipopolysaccharide (LPS)-induced expression of the reporter (luciferase) gene, although the proximal κB site played the more essential role. Electrophoretic mobility shift assay revealed that LPS augmented the nuclear protein bindings to the Ets/PEA3 as well as κB motifs. Ets-1, one of the Ets proteins, was suggested to be bound to the Ets/PEA3 oligonucleotide. By Northern blot analysis, LPS was shown to induce iNOS mRNA in CEVM, along with a preceding increase in the levels of c-ets-1 mRNA. Ets-1 may be involved in the iNOS gene transcription in CEVM, presumably through interacting with the NF-κB.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Crepieux P, Coll J, Stehelin D: The Ets family of proteins: Weak modulators of gene expression in quest for transcriptional partners. Crit Rev Oncogenesis 5: 615–638, 1994

    Google Scholar 

  2. Gutman A, Wasylyk B: The collagenase gene promoter contains a TPA and oncogene-responsive unit encompassing the PEA3 and AP-1 binding sites. EMBO J 9: 2241–2246, 1990

    Google Scholar 

  3. Krämer B, Wiegmann K, Kronke M: Regulation of the human TNF promoter by the transcription factor Ets. J Biol Chem 270: 6577–6583, 1995

    Google Scholar 

  4. Bassuk AG, Anandappa RT, Leiden JM: Physical interaction between Ets and NF-κB/NFAT proteins plays an important role in their cooperative activation of the human immunodeficiency virus enhancer in T cells. J Virol 71: 3563–3573, 1997

    Google Scholar 

  5. Gri G, Savio D, Trinchieri G, Ma X: Synergistic regulation of the human interleukin-12 p40 promoter by NFκB and Ets transcription factors in Epstein-Barr virus-transformed B cells and macrophages. J Biol Chem 273: 6431–6438, 1998

    Google Scholar 

  6. Wasylyk B, Wasylyk C, Flores P, Begue A, Leprince D, Stehelin D: The c-ets proto-oncogenes encodes transcription factors that cooperate with c-Fos and c-Jun for transcriptional activation. Nature 346: 191–193, 1990

    Google Scholar 

  7. Cieslik K, Zembowicz A, Tang J-L, Wu KK: Transcriptional regulation of endothelial nitric-oxide synthase by lysophosphatidylcholine. J Biol Chem 273: 14885–14890, 1998

    Google Scholar 

  8. Kelly RA, Balligand J-L, Smith TW: Nitric oxide and cardiac function. Circ Res 79: 363–380, 1996

    Google Scholar 

  9. de Belder AJ, Radomski MW, Why HJF, Richardson PJ, Bucknall CA, Salas E, Martin JF, Moncada S: Nitric oxide synthase activities in human myocardium. Lancet 341: 84–85, 1993

    Google Scholar 

  10. de Belder AJ, Radomski MW, Why HJF, Richardson PJ, Martin JF: Myocardial calcium-independent nitric oxide synthase activity is present in dilated cardiomyopathy, myocarditis, and postpartum cardiomyopathy but not in ischemic or valvular heart disease. Br Heart J 74: 426–430, 1995

    Google Scholar 

  11. Habib FM, Springall DR, Davies GJ, Oakley CM, Yacoub MH, Polak JM: Tumor necrosis factor and inducible nitric oxide synthase in dilated cardiomyopathy. Lancet 347: 1151–1155, 1996

    Google Scholar 

  12. Lewis NP, Tsao PS, Rickenbacher PR, Xue C, Johns RA, Haywood GA, von der Leyen H, Trindale PT, Cooke JP, Hunt SA, Billingham ME, Valantine HA, Fowler MB: Induction of nitric oxide synthase in the human cardiac allograft is associated with contractile dysfunction of the left ventricle. Circulation 93: 720–729, 1996

    Google Scholar 

  13. Haywood GA, Tsao PS, von der Leyen HE, Mann MJ, Keeling PJ, Trindale PT, Lewis NP, Byrne CD, Rickenbacher PR, Bishopric NH, Cooke JP, McKenna WJ, Fowler MB: Expression of inducible nitric oxide synthase in human heart failure. Circulation 93: 1087–1094, 1996

    Google Scholar 

  14. Satoh M, Nakamura M, Tamura G, Makita S, Segawa I, Tashiro A, Satodate R, Hiramori K: Inducible nitric oxide synthase and tumor necrosis factor-alpha in myocardium in human dilated cardiomyopathy. J Am Coll Cardiol 29: 716–724, 1997

    Google Scholar 

  15. Fukuchi M, Hussain SNA, Giaid A: Heterogeneous expression and activity of endothelial and inducible nitric oxide synthases in end-stage human heart failure: Their relation to lesion site and β-adrenergic receptor therapy. Circulation 98: 132–139, 1998

    Google Scholar 

  16. Kinugawa K, Shimizu T, Yao A, Kohmoto O, Serizawa T, Takahashi T: Transcriptional regulation of inducible nitric oxide synthase in cultured neonatal rat cardiac myocytes. Circ Res 81: 911–921, 1997

    Google Scholar 

  17. Lowenstein CJ, Alley EW, Raval P, Snowman AM, Snyder SH, Russel SW, Murphy WJ: Macrophage nitric oxide synthase gene: Two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc Natl Acad Sci USA 90: 9730–9734, 1993

    Google Scholar 

  18. Xie Q-W, Whisnant R, Nathan C: Promoters of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon γ and bacterial lipopolysaccharide. J Exp Med 177: 1779–1784, 1993

    Google Scholar 

  19. Eberhardt W, Kunz D, Hummel R, Pfeilschifter J: Molecular cloning of the rat inducible nitric oxide synthase gene promoter. Biochem Biophys Res Commun 223: 752–756, 1996

    Google Scholar 

  20. de Vera ME, Shapiro RA, Nussler AK, Mudget JS, Simmons RL. Morris SM Jr, Billar TR, Geller DA: Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: Initial analysis of the human NOS2 promoter. Proc Natl Acad Sci USA 93: 1054–1059, 1996

    Google Scholar 

  21. Lin AW, Chang CC, McCormick CC: Molecular cloning and expression of an avian macrophage nitric-oxide synthase cDNA and the analysis of genomic 5′-flanking region. J Biol Chem 271: 11911–11919, 1996

    Google Scholar 

  22. Taylor BS, de Vera ME, Ganster RW, Wang Q, Shapiro RA, Morris SM Jr, Billiar TR, Gellar DA: Multiple NF-κB enhancer elements regulate cytokine induction of the human inducible nitric oxide synthase gene. J Biol Chem 273: 15148–15156, 1998

    Google Scholar 

  23. Marks-Konczalik J, Chu SC, Moss J: Cyotokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor κB-binding sites. J Biol Chem 273: 22201–22208, 1998

    Google Scholar 

  24. Shimizu T, Kinugawa K, Sugishita Y, Sugishita K, Harada K, Matsui H, Kohmoto O, Serizawa T, Takahashi T: Molecular cloning and expression of inducible nitric oxide synthase in chick embryonic ventricular myocytes. Cardiovasc Res 38: 405–413, 1998

    Google Scholar 

  25. Schreiber E, Matthias P, Müller MM, Schaffner W: Rapid detection of octamer binding proteins with ‘mini-extracts', prepared from a small number of cells. Nucleic Acids Res 17: 6419, 1989

    Google Scholar 

  26. Leprince D, Duterque-Coquillaud M, Li R-P, Henry C, Flourens A, Debiure B, Stehelin D: Alternative splicing within the chicken c-ets-1 locus: Implications for transduction within the E26 retrovirus of the c-ets proto-oncogene. J Virol 62: 3233–3241, 1988

    Google Scholar 

  27. Wang LG, Liu XM, Li ZR, Denstman S, Bloch A: Differential binding of nuclear c-ets-1 protein to an intron I fragment of the c-myb gene in growth vs. differentiation. Cell Growth Diff 5: 1243–1251, 1994

    Google Scholar 

  28. Ko JH, Miyoshi E, Noda K, Ekuni A, Kang R, Ikeda Y, Taniguchi N: Regulation of the GnT-V promoter by transcription factor Ets-1 in various cancer cell lines. J Biol Chem 274: 22941–22948, 1999

    Google Scholar 

  29. Shimizu T, Kinugawa K, Yao A, Sugishita Y, Sugishita K, Harada K, Matsui H, Kohmoto O, Serizawa T, Takahashi T: Platelet-derived growth factor induces cellular growth in cultured chick ventricular myocytes. Cardiovasc Res 41: 641–653, 1999

    Google Scholar 

  30. Strickler J, Jacobson KA, Liang BT: Direct preconditioning of cultured chick ventricular myocytes. Novel functions of cardiac adenosine A2a and A3 receptors. J Clin Invest 98: 1773–1779, 1996

    Google Scholar 

  31. Bhat NK, Thompson CB, Lindsten T, June CH, Fujiwara S, Koizumi S, Fisher RJ, Papas TS: Reciprocal expression of human ETS1 and ETS2 genes during T-cell activation: Regulatory role for the protooncogene ETS1. Proc Natl Acad Sci USA 87: 3723–3727, 1990

    Google Scholar 

  32. Maroulakou IG, Papas TS, Green JE: Differential expression of ets-1 and ets-2 proto-oncogenes during murine embryogenesis. Oncogene 9: 1551–1565, 1994

    Google Scholar 

  33. Baeuerle PA, Baltimore D: NF-κB: Ten years after. Cell 87: 13–20, 1996

    Google Scholar 

  34. Baldwin AS Jr: The NF-κB and IκB proteins: New discoveries and insights. Annu Rev Immunol 14: 649–681, 1996

    Google Scholar 

  35. Xie Q-W, Kashiwabara Y, Nathan C: Role of transcription factor NFkappaB/ Rel in induction of nitric oxide synthase. J Biol Chem 269: 4705–4708, 1994

    Google Scholar 

  36. Oddis CV, Finkel MS: NF-κB and GTP cyclohydrolase regulate cytokine-induced nitric oxide production by cardiac myocytes. Am J Physiol 270: H1864–H1868, 1996

    Google Scholar 

  37. Peterson PK, Hu S, Anderson WR, Chao CC: Nitric oxide production and neurotoxicity mediated by activated microglia from human vs. mouse brain. J Infect Dis 170: 457–460, 1994

    Google Scholar 

  38. Weinberg JB, Misukonis MA, Shami PJ, Mason SN, Sauls DL, Dittman WA, Wood ER, Smith GK, McDonald B, Bachus KE, Haney AF, Granger DL: Human mononuclear phagocyte inducible nitric oxide synthase (iNOS): Analysis of iNOS mRNA, iNOS protein, biopterin, and nitric oxide production by blood monocyte and peritoneal macrophage. Blood 86: 1184–1195, 1995

    Google Scholar 

  39. Adler H, Frech B, Thony M, Pfister H, Peterhans E, Jungi TW: Inducible nitric oxide synthase in cattle. Differential cytokine regulation of nitric oxide synthase in bovine and murine macrophages. J Immunol 154: 4710–4718, 1995

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Toshiyuki Takahashi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takahashi, T., Sugishita, Y., Kinugawa, Ki. et al. Ets-1 is involved in transcriptional regulation of the chick inducible nitric oxide synthase gene in embryonic ventricular myoctyes. Mol Cell Biochem 226, 57–65 (2001). https://doi.org/10.1023/A:1012781618109

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1012781618109

Navigation