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Thromb Haemost 2000; 84(06): 1031-1038
DOI: 10.1055/s-0037-1614167
Review Article
Schattauer GmbH

Characterization of the Murine Coagulation Factor X Promoter

Julie A. Wilberding
1   From the Walther Cancer Center, the W. M. Keck Center for Transgene Research, and the Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA
,
Francis J. Castellino
1   From the Walther Cancer Center, the W. M. Keck Center for Transgene Research, and the Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA
› Author AffiliationsWe thank Mr. Ivo Cornelissen for his assistance with tissue removal from mice. This work was supported in part by grants HL19982 from the N.I.H. (to FJC), the Kleiderer-Pezold endowed professorship (to FJC), and a grant from the W. M. Keck Foundation. These studies were performed in collaboration with the Walther Cancer Foundation, Indianapolis, IN.
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Publication History

Received 18 April 2000

Accepted after revision 14 July 2000

Publication Date:
13 December 2017 (online)

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Summary

Factor X (FX) is a vitamin K-dependent serine protease zymogen that functions in both the extrinsic and intrinsic pathways of blood coagulation. In this study, the 5’-flanking region of the murine FX gene was analyzed to determine those elements that govern its transcriptional activity and regulation. Consistent with other TATA-less promoters, murine FX contains two start sites of transcription, at bp −5 and −21 relative to the ATG translational initiation codon. The mRNA of FX was found in a number of tissues, including the liver, stomach, intestine, kidney, ovary, testes, spleen, skeletal muscle, and lung. Using DNase I footprinting, three areas of protection have been identified in the proximal 287 bp of the promoter, spanning bp −28 to −218. Further examination of this region revealed transcription factor binding sites for NF-Y, HNF-4, and a GATA factor. Electrophoretic mobility shift analysis (EMSA) confirmed the identities of NF-Y, HNF-4, and GATA-4, all of which were found by transient transfection analyses in HepG2 cells to influence the activity of the promoter. Ablation of the NF-Y site was most dramatic, reducing activity to 10% of that of the wild-type construct. Deletion of the HNF-4 site led to an activity of 25% of wild-type, and a GATA-4 mutation reduced activity to 63% of wild-type values. This investigation revealed the identity of the factors bound at the proximal promoter of the FX gene, and the relative importance of each. This is the first report of a member of the GATA family of transcription factors being important in the regulation of a coagulation-based gene.

Key words

Factor X - promoter analysis - gene regulation - transcription factors - coagulation
 

  • References

  • 1 Jackson CM, Nemerson Y. Blood coagulation. Ann Rev Biochem 1980; 49: 765-811.
    CrossrefPubMedGoogle Scholar
  • 2 Baugh RJ, Broze GJ, Krishnaswamy S. Regulation of extrinsic pathway factor Xa formation by tissue factor pathway inhibitor. J Biol Chem 1998; 273: 4378-86.
    CrossrefPubMedGoogle Scholar
  • 3 Broze GJ. Tissue factor pathway inhibitor and the current concept of blood coagulation. Blood Coagul Fibrinolysis 1995; 06 (Suppl. 01) S7-13.
    CrossrefPubMedGoogle Scholar
  • 4 McMullen BA, Fujikawa K, Kisiel W, Sasagawa T, Howald WN, Kwa EW, Weinstein B. Complete amino acid sequence of the light chain of human blood coagulation factor X: Evidence for identification of residue 63 as β-hydroxyaspartic acid. Biochemistry 1983; 22: 2875-84.
    CrossrefPubMedGoogle Scholar
  • 5 Leytus SP, Chung DW, Kisiel W, Kurachi K, Davie EW. Characterization of a cDNA coding for human factor X. Proc Natl Acad Sci USA 1984; 81: 3699-702.
    CrossrefPubMedGoogle Scholar
  • 6 Leytus SP, Foster DC, Kurachi K, Davie EW. Gene for human factor X: A blood coagulation factor whose gene organization is essentially identical with that of factor IX and protein C. Biochemistry 1986; 25: 5098-102.
    CrossrefPubMedGoogle Scholar
  • 7 Liang Z, Cooper A, DeFord ME, Carmeliet P, Collen D, Castellino FJ, Rosen ED. Cloning and characterization of a cDNA encoding murine coagulation factor X. Thromb Haemost 1998; 80: 87-91.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Cooper A, Liang Z, Castellino FJ, Rosen ED. Cloning and characterization of the murine coagulation factor X gene. Thromb Haemost 2000; 83: 732-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Gasic GP, Arenas CP, Gasic TB, Gasic GJ. Coagulation factors X, Xa, and protein S as potent mitogens of cultured aortic smooth muscle cells. Proc Natl Acad Sci USA 1992; 89: 2317-20.
    CrossrefPubMedGoogle Scholar
  • 10 Cirino G, Cicala C, Bucci M, Sorrentino L, Ambrosini G. Factor Xa as an interface between coagulation and inflammation. Molecular mimicry of factor Xa association with effector cell protease receptor-1 induces acute inflammation in vivo. J Clin Invest 1997; 99: 2446-51.
    CrossrefPubMedGoogle Scholar
  • 11 Dewerchin M, Liang Z, Moons L, Carmeliet P, Castellino FJ, Collen D, Rosen ED. Blood coagulation factor X deficiency causes partial embryonic lethality and fatal neonatal bleeding in mice. Thromb Haemost 2000; 83: 185-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Liang Z, Cooper A, Rosen ED, Castellino FJ. Chromosomal arrangement of the murine coagulation factor VII and factor X genes. Thromb Haemost 1998; 80: 524-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Kornberg RD, LaPointe JW, Lorch Y. Preparation of nucleosomes and chromatin. Methods Enzymol 1989; 170: 3-14.
    PubMedGoogle Scholar
  • 14 Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 1983; 11: 1475-89.
    CrossrefPubMedGoogle Scholar
  • 15 Stauffer DR, Chukwumezie BN, Wilberding JA, Rosen ED, Castellino FJ. Characterization of transcriptional regulatory elements in the promoter region of the murine blood coagulation factor VII gene. J Biol Chem 1998; 273: 2277-87.
    CrossrefPubMedGoogle Scholar
  • 16 Huang N-M, Hung H-L, Stanfield-Oakley SA, High KA. Characterization of the human blood coagulation factor X promoter. J Biol Chem 1992; 267: 15440-8.
    PubMedGoogle Scholar
  • 17 Hung HL, High KA. Liver enriched transcription factor HNF-4 and ubiquitous factor NF-Y are critical for expression of blood coagulation factor X. J Biol Chem 1996; 271: 2323-31.
    CrossrefPubMedGoogle Scholar
  • 18 Quandt K, Frech K, Karas H, Wingender E, Werner T. MatInd and Mat-Inspector: new fast versatile tools for detection of consensus matches in nucleotide sequence data. Nucl Acids Res 1995; 23: 4878-84.
    CrossrefPubMedGoogle Scholar
  • 19 Bossard P, Zaret KS. GATA transcription factors as potentiators of gut endoderm differentiation. Development 1998; 125: 4909-17.
    PubMedGoogle Scholar
  • 20 Miao CH, Leytus SP, Chung DW, Davie EW. Liver-specific expression of the gene coding for human factor X, a blood coagulation factor. J Biol Chem 1992; 267: 7395-401.
    PubMedGoogle Scholar
  • 21 Reijnen MJ, Sladek FM, Bertina RM, Reitsma PH. Disruption of a binding site for hepatocyte nuclear factor 4 results in hemophilia B Leyden. Proc Natl Acad Sci USA 1992; 89: 6300-3.
    CrossrefPubMedGoogle Scholar
  • 22 Blake MC, Jambou RC, Swick AG, Kahn JW, Azizkhan JC. Transcriptional initiation is controlled by upstream GC-box interactions in a TATAAless promoter. Mol Cell Biol 1990; 10: 6632-41.
    CrossrefPubMedGoogle Scholar
  • 23 Geng Y, Johnson LF. Lack of an initiator element is responsible for multiple transcriptional initiation sites of the TATA-less mouse thymidylate synthase promoter. Mol Cell Biol 1993; 13: 4894-903.
    CrossrefPubMedGoogle Scholar
  • 24 McGlynn LK, Mueller CR, Begbie M, Notley CR, Lillicrap D. Role of the liver-enriched transcription factor hepatocyte nuclear factor 1 in transcriptional regulation of the factor VIII gene. Mol Cell Biol 1996; 16: 1936-45.
    CrossrefPubMedGoogle Scholar
  • 25 Greenberg D, Miao CH, Ho WT, Chung DW, Davie EW. Liver-specific expression of the human factor VII gene. Proc Natl Acad Sci USA 1995; 92: 12347-51.
    CrossrefPubMedGoogle Scholar
  • 26 Pollak ES, Hung HL, Godin W, Overton GC, High KA. Functional characterization of the human factor VII 5’-flanking region. J Biol Chem 1996; 271: 1738-47.
    CrossrefPubMedGoogle Scholar
  • 27 Reijnen MJ, Bertina RM, Reitsma PH. Localization of transcription initiation sites in the human coagulation factor IX gene. FEBS Lett 1990; 270: 207-10.
    CrossrefPubMedGoogle Scholar
  • 28 Dorn A, Bollekens J, Staub A, Benoist C, Mathis D. A multiplicity of CCAAT box-binding proteins. Cell 1987; 50: 863-72.
    CrossrefPubMedGoogle Scholar
  • 29 Berry M, Grosveld F, Dillon N. A single point mutation is the cause of the Greek form of hereditary persistence of fetal haemoglobin. Nature 1992; 358: 499-502.
    CrossrefPubMedGoogle Scholar
  • 30 Ronchi A, Berry M, Raguz S, Imam A, Yannoutsos N, Ottolenghi S, Grosveld F, Dillon N. Role of the duplicated CCAAT box region in gammaglobin gene regulation and hereditary persistence of fetal haemoglobin. EMBO J 1996; 15: 143-9.
    PubMedGoogle Scholar
  • 31 Roy B, Lee AS. Transduction of calcium stress through interaction of the human transcription factor CBF with the proximal CCAAT regulatory element of the grp78/BiP promoter. Mol Cell Biol 1995; 15: 2263-74.
    CrossrefPubMedGoogle Scholar
  • 32 Marziali G, Perrotti E, Ilari R, Testa U, Coccia EM, Battistini A. Transcriptional regulation of the ferritin heavy-chain gene: the activity of the CCAAT binding factor NF-Y is modulated in heme-treated Friend leukemia cells and during monocyte-to-macrophage differentiation. Mol Cell Biol 1997; 17: 1387-5.
    CrossrefPubMedGoogle Scholar
  • 33 Mantovani R. The molecular biology of the CCAAT-binding factor NF-Y. Gene 1999; 239: 15-27.
    CrossrefPubMedGoogle Scholar
  • 34 Jackson SM, Ericsson J, Mantovani R, Edwards PA. Synergistic activation of transcription by nuclear factor Y and sterol regulatory element binding protein. J Lipid Res 1998; 39: 767-76.
    PubMedGoogle Scholar
  • 35 Wright KL, Moore TL, Vilen BJ, Brown AM, Ting JP. Major histocompatibility complex class II-associated invariant chain gene expression is upregulated by cooperative interactions of Sp1 and NF-Y. J Biol Chem 1995; 270: 20978-86.
    CrossrefPubMedGoogle Scholar
  • 36 Ueda A, Takeshita F, Yamashiro S, Yoshimura T. Positive regulation of the human macrophage stimulating protein gene transcription. Identification of a new hepatocyte nuclear factor-4 (HNF-4) binding element and evidence that indicates direct association between NF-Y and HNF-4. J Biol Chem 1998; 273: 19339-47.
    CrossrefPubMedGoogle Scholar
  • 37 Sladek FM, Zhong WM, Lai E, Darnell Jr JE. Liver-enriched transcription factor HNF-4 is a novel member of the steroid hormone receptor superfamily. Genes Dev 1990; 04: 2353-65.
    CrossrefPubMedGoogle Scholar
  • 38 Costa RH, Grayson DR, Darnell JEJ. Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol Cell Biol 1989; 09: 1415-25.
    CrossrefPubMedGoogle Scholar
  • 39 Duncan SAKM, Chen WS, Hoodless P, Weinstein DC, Bachvarova RF, Darnell Jr JE. Expression of transcription factor HNF-4 in the extra- embryonic endoderm, gut, and nephrogenic tissue of the developing mouse embryo: HNF-4 is a marker for primary endoderm in the implanting blastocyst. Proc Natl Acad Sci USA 1994; 91: 7598-602.
    CrossrefPubMedGoogle Scholar
  • 40 Taraviras S, Monaghan AP, Schutz G, Kelsey G. Characterization of the mouse HNF-4 gene and its expression during mouse embryogenesis. Mech Dev 1994; 48: 67-79.
    CrossrefPubMedGoogle Scholar
  • 41 Cereghini S. Liver-enriched transcription factors and hepatocyte differentiation. FASEB J 1996; 10: 267-82.
    CrossrefPubMedGoogle Scholar
  • 42 Metzger S, Halaas JL, Breslow JL, Sladek FM. Orphan receptor HNF-4 and bZip protein C/EBP alpha bind to overlapping regions of the apolipoprotein B gene promoter and synergistically activate transcription. J Biol Chem 1993; 268: 16831-8.
    PubMedGoogle Scholar
  • 43 Laverriere AC, MacNeill C, Mueller C, Poelmann RE, Burch JB, Evans T. GATA-4/5/6, a subfamily of three transcription factors transcribed in developing heart and gut. J Biol Chem 1994; 269: 23177-84.
    PubMedGoogle Scholar