Identification of a Glucocorticoid Response Element in the Human γ Chain Fibrinogen Promoter

Summary

The effect of the synthetic glucocorticoid hormone dexamethasone on human γ chain fibrinogen gene expression was examined. The whole promoter region of 3.8 kb of this gene and progressive 5’-deletions were inserted into a promoterless expression vector, upstream of the luciferase gene and transiently transfected into the human hepatoma HepG2 cells, in the presence or in the absence of dexamethasone stimulation. Deletion analysis allowed to identify a region located between –1359 and –954 bp upstream from the transcription start site, involved in hormone inducibility. On the basis of a computer-assisted analysis, a putative GRE was found in this region at bases –1116 to –1102. Specific point mutations eliminating this putative GRE led to complete loss of glucocorticoid inducibility, thus indicating its functional role. Binding of the rat glucocorticoid receptor to this site was demonstrated by mobility-shift assays.

• References

• 1 Doolittle RF. Fibrinogen and fibrin. Ann Rev Biochem 1984; 53: 195-229.
• 2 Marino MW, Fuller GM, Elder FFB. Chromosomal localization of human and rat Aα, Bβ, and γ fibrinogen genes by in situ hybridization. Cytogen Cell Genet 1986; 42: 36-41.
• 3 Kant JA, Fornace Jr AJ, Save D, Simon MI, McBride OW, Crabtree GR. Evolution and organization of the fibrinogen locus on chromosome 4: gene duplication accompanied by transposition and inversion. Proc Natl Acad Sci USA 1985; 82: 2344-8.
• 4 Roy S, Overton O, Redman C. Overexpression of any fibrinogen chain by HepG2 cells specifically elevates the expression of the other two chains. J Biol Chem 1994; 269: 691-5.
• 5 Roberts LR, Holland LJ. Coordinate transcriptional regulation of the three fibrinogen subunit genes by glucocorticoids in cultured primary liver cells from Xenopus laevis. Endocrinology 1993; 132: 2563-70.
• 6 Blombäck B. Fibrinogen structure, activation, polymerization and fibrin gel structure. Thromb Res 1994; 75: 327-8.
• 7 Hettasch JM, Bolyard MG, Lord ST. The residues AGDV of recombinant γ chains of human fibrinogen must be carboxy-terminal to support human platelet aggregation. Thromb Haemost 1992; 68: 701-6.
• 8 De la Serna G. Fibrinogen: a new major risk factor for cardiovascular disease. A review of the literature. J Fam Pract 1994; 39: 468-77.
• 9 Mannucci PM, Mari D. Fibrinogen as a model protein in epidemiological studies. Fibrinolysis 1993; 7: 51-2.
• 10 Kushner I. The acute-phase response: an overview. Methods Enzymol 1988; 163: 373-83.
• 11 Higazi AA, Barghouti II, Ayesh S, Mayer M, Matzner Y. Inhibition of neutrophil activation by fibrinogen. Inflammation 1994; 18: 525-35.
• 12 Baumann H, Prowse KR, Marinkovic S, Won KA, Jahries GP. Stimulation of hepatic acute phase response by cytokines and glucocorticoids. Ann NY Acad Sci 1989; 557: 280-95.
• 13 Van Snick J. Interleukin-6: an overview. Ann Rev Immunol 1990; 8: 253-78.
• 14 Huber P, Laurent M, Dalmon J. Human β-fibrinogen gene expression: upstream sequences involved in its tissue specific expression and its dexamethasone and interleukin-6 stimulation. J Biol Chem 1990; 265: 5695-701.
• 15 Anderson GM, Shaw AR, Shafer JA. Functional characterization of promoter elements involved in regulation of human Bβ fibrinogen expression: evidence for binding of novel activator and repressor proteins. J Biol Chem 1993; 268: 22650-5.
• 16 Dalmon J, Laurent M, Courtois G. The human β fibrinogen promoter contains a hepatocyte nuclear factor 1-dependent interleukin-6-responsive element. Mol Cell Biol 1993; 13: 1183-93.
• 17 Hu CH, Harris JE, Davie EW, Chung DW. Characterization of the 5’-flanking region of the gene for the α chain of human fibrinogen. J Biol Chem 1995; 270: 28342-9.
• 18 Mizuguchi J, Hu CH, Cao Z, Loeb KR, Chung DW, Davie EW. Characterization of the 5’-flanking region of the gene for the α chain of human fibrinogen. J Biol Chem 1995; 270: 28350-6.
• 19 Baumann H, Jahreis GP, Morella KK. Interaction of cytokine- and glucocorticoid-response elements of acute-phase plasma protein and genes. Importance of glucocorticoid receptor level and cell type for regulation of the elements from rat α₁-acid glycoprotein and β fibrinogen genes. J Biol Chem 1990; 265: 22275-81.
• 20 Liu Z, Fuller GM. Detection of a novel transcription factor for the Aα fibrinogen gene in response to interleukin-6. J Biol Chem 1995; 270: 7580-6.
• 21 Zhang Z, Fuentes NL, Fuller GM. Characterization of the IL-6 responsive elements in the γ fibrinogen gene promoter. J Biol Chem 1995; 270: 24287-91.
• 22 Zhong Z, Wen Z, Darnell Jr. JE. Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. Science 1994; 264: 95-8.
• 23 Marchetti L, Zanelli T, Malcovati M, Tenchini ML. Polymorphism of the human gamma chain fibrinogen gene. DNA Seq 1991; 1: 419-22.
• 24 Rixon MW, Chung DW, Davie EW. Nucleotide sequence of the gene for the γ chain of human fibrinogen. Biochemistry 1985; 24: 2077-86.
• 25 Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. Cold Spring Harbor Laboratory; Cold Spring Harbor, NY: 1989
• 26 Wigler M, Pellicer A, Silverstein S, Axel R. Biochemical transfer of single-copy eukaryotic gene using total cellular DNA as donor. Cell 1978; 14: 725-31.
• 27 Luisi BF, Xu WX, Otwinowski Z, Freedman LP, Yamamoto KR, Sigler PB. Crystallographic analysis of the interaction of the glucocorticoid receptor with DNA. Nature 1991; 352: 478-9.
• 28 Roberts LR, Nichols LA, Holland LJ. Transcriptional regulation of the Xenopus laevis B β fibrinogen subunit gene by glucocorticoids and hepatocyte nuclear factor 1: analysis by transfection into primary liver cells. Biochemistry 1993; 32: 11627-37.
• 29 Beato M. Gene regulation by steroid hormones. Cell 1989; 56: 335-44.
• 30 Faisst S, Meyer S. Compilation of vertebrate-encoded transcription factors. Nucleic Acids Res 1992; 20: 3-26.
• 31 La Baer J, Yamamoto KR. Analysis of the DNA-binding affinity, sequence specificity and context dependence of the glucocorticoid receptor zinc finger region. J Mol Biol 1994; 239: 664-88.
• 32 Tsai SY, Carlstedt-Duke J, Weiglel NL, Dahlman K, Gustafsson JA, Tsai MJ, O’Malley BW. Molecular interactions of steroid hormone receptor with its enhancer element: evidence for receptor dimer formation. Cell 1988; 55: 361-9.
• 33 Lucas PC, Granner DK. Hormone response domains in gene transcription. Ann Rev Biochem 1992; 61: 1131-73.
• 34 Woodward RN, Li M, Holland LJ. Novel accessory factor-binding site required for glucocorticoid regulation of the gamma-fibrinogen subunit gene from Xenopus laevis. Mol Endocrinol 1997; 11: 563-76.
• 35 Buetti E, Kühnel B. Distinct sequence elements involved in the glucocorticoid regulation of the mouse mammary tumor virus promoter identified by linker scanning mutagenesis. J Mol Biol 1986; 190: 379-89.
• 36 Raju VS, McCoubrey Jr WK, Maines MD. Regulation of heme oxygenase-2 by glucocorticoids in neonatal rat brain: characterization of a functional glucocorticoid response element. Biochim Biophys Acta 1997; 1351: 89-104.
• 37 Janzten HM, Strahle U, Gloss B, Stewart F, Schmid W, Boshart M, Miksicek R, Schutz G. Cooperativity of glucocorticoid response elements located far upstream of the tyrosine aminotransferare gene. Cell 1987; 49: 29-38.
• 38 Di Lorenzo D, Williams P, Ringold G. Identification of two distinct nuclear factors with DNA binding activity within the glucocorticoid regulatory region of the rat α₁-acid glycoprotein promoter. Biochem Biophys Res Commun 1991; 176: 1326-32.
• 39 Karin M, Haslinger A, Holtgreve H, Richards RI, Krauter P, Westphal HM, Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature 1984; 308: 513-9.
• 40 Argenton F, Ramoz N, Charlet N, Bernardini S, Colombo L, Bortolussi M. Mechanisms of transcriptional activation of the promoter of the rainbow trout prolactin gene by GHF1/Pit1 and glucocorticoid. Biochem Biophys Res Commun 1996; 224: 57-66.
• 41 Butz K, Hoppe-Seyler F. Transcriptional control of human papillomavirus (HPV) oncogene expression: composition of the HPV type 18 upstream regulatory region. J Virol 1993; 67: 6476-86.
• 42 Costa RH, Grayson DR, Darnell Jr. JE. Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol Cell Biol 1989; 9: 1415-25.
• 43 O’Brien RM, Noisin EL, Suwanichkul A, Yamasaki T, Lucas PC, Wang JC, Powell DR, Granner DK. Hepatic nuclear factor 3- and hormone-regulated expression of the phosphoenolpyruvate carboxykinase and insulin-like growth factor-binding protein 1 genes. Mol Cell Biol 1995; 15: 1747-58.
• 44 Unterman TG, Fareeduddin A, Harris MA, Goswami RG, Porcella A, Costa RH, Lacson RG. Hepatocyte nuclear factor-3 (HNF-3) binds to the insulin response sequence in the IGF binding protein-1 (IGFBP-1) promoter and enhances promoter function. Biochem Biophys Res Commun 1994; 203: 1835-41.
• 45 Nitsch D, Boshart M, Schutz G. Activation of the tyrosine aminotransferase gene is dependent on synergy between liver-specific and hormone-responsive elements. Proc Natl Acad Sci USA 1993; 90: 5479-83.