Skip to main content
PMC home page
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
. 1992 Jul 15;89(14):6300–6303. doi: 10.1073/pnas.89.14.6300

Disruption of a binding site for hepatocyte nuclear factor 4 results in hemophilia B Leyden.

M J Reijnen 1, F M Sladek 1, R M Bertina 1, P H Reitsma 1
PMCID: PMC49488  PMID: 1631121

Abstract

Hemophilia B Leyden is an X chromosome-linked bleeding disorder characterized by very low plasma levels of blood coagulation factor IX (fIX) during childhood. After puberty, plasma fIX levels gradually rise to a maximum of 60% of normal, probably under the influence of testosterone. Single point mutations in the fIX promoter region of hemophilia B Leyden patients have been reported at -20, -6, -5, +8 and +13. In addition, one promoter mutation (G----C at -26) has been detected that abolishes fIX expression throughout life (M. Ludwig, personal communication). We examined how one of the hemophilia B Leyden mutations (T----A at -20) and the G----C mutation at -26 interfere with fIX gene transcription. We report that the wild-type promoter of the human fIX gene contains a binding site (at nucleotides -34 to -10) for hepatocyte nuclear factor 4 (HNF-4), a member of the steroid hormone receptor superfamily of transcription factors. The binding of HNF-4 is disrupted by both the T----A mutation at -20 and the G----C mutation at -26. Whereas HNF-4 transactivates the wild-type promoter sequence in liver (HepG2) and non-liver (HeLa) cell types quite well, it transactivates the -20 mutated promoter to only a limited extent and the -26 mutated promoter not at all. These data suggest that HNF-4 is a major factor controlling fIX expression in the normal individual and that its inability to bind efficiently to the -20 T----A and the -26 G----C mutated promoter sequence results in hemophilia. Further, the severity of the hemophilia phenotype appears to be directly related to the degree of disruption of HNF-4 binding and transactivation.

Full text

PDF
6300

Images in this article

6302Image
on p.6302
6302Image
on p.6302

Selected References

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

  1. Briët E., Bertina R. M., van Tilburg N. H., Veltkamp J. J. Hemophilia B Leyden: a sex-linked hereditary disorder that improves after puberty. N Engl J Med. 1982 Apr 1;306(13):788–790. doi: 10.1056/NEJM198204013061306. [DOI] [PubMed] [Google Scholar]
  2. Briët E., Wijnands M. C., Veltkamp J. J. The prophylactic treatment of hemophilia B Leyden with anabolic steroids. Ann Intern Med. 1985 Aug;103(2):225–226. doi: 10.7326/0003-4819-103-2-225. [DOI] [PubMed] [Google Scholar]
  3. Costa R. H., Grayson D. R., Darnell J. E., Jr Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol Cell Biol. 1989 Apr;9(4):1415–1425. doi: 10.1128/mcb.9.4.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crossley M., Brownlee G. G. Disruption of a C/EBP binding site in the factor IX promoter is associated with haemophilia B. Nature. 1990 May 31;345(6274):444–446. doi: 10.1038/345444a0. [DOI] [PubMed] [Google Scholar]
  5. Edlund T., Walker M. D., Barr P. J., Rutter W. J. Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5' flanking elements. Science. 1985 Nov 22;230(4728):912–916. doi: 10.1126/science.3904002. [DOI] [PubMed] [Google Scholar]
  6. Evans R. M. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. doi: 10.1126/science.3283939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Forman B. M., Samuels H. H. Dimerization among nuclear hormone receptors. New Biol. 1990 Jul;2(7):587–594. [PubMed] [Google Scholar]
  8. Giannelli F., Green P. M., High K. A., Sommer S., Lillicrap D. P., Ludwig M., Olek K., Reitsma P. H., Goossens M., Yoshioka A. Haemophilia B: database of point mutations and short additions and deletions--second edition. Nucleic Acids Res. 1991 Apr 25;19 (Suppl):2193–2219. doi: 10.1093/nar/19.suppl.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glass C. K., Lipkin S. M., Devary O. V., Rosenfeld M. G. Positive and negative regulation of gene transcription by a retinoic acid-thyroid hormone receptor heterodimer. Cell. 1989 Nov 17;59(4):697–708. doi: 10.1016/0092-8674(89)90016-0. [DOI] [PubMed] [Google Scholar]
  10. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  11. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  12. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  13. O'Malley B. The steroid receptor superfamily: more excitement predicted for the future. Mol Endocrinol. 1990 Mar;4(3):363–369. doi: 10.1210/mend-4-3-363. [DOI] [PubMed] [Google Scholar]
  14. Reijnen M. J., Bertina R. M., Reitsma P. H. Localization of transcription initiation sites in the human coagulation factor IX gene. FEBS Lett. 1990 Sep 17;270(1-2):207–210. doi: 10.1016/0014-5793(90)81269-t. [DOI] [PubMed] [Google Scholar]
  15. Rosenfeld P. J., Kelly T. J. Purification of nuclear factor I by DNA recognition site affinity chromatography. J Biol Chem. 1986 Jan 25;261(3):1398–1408. [PubMed] [Google Scholar]
  16. Salier J. P., Hirosawa S., Kurachi K. Functional characterization of the 5'-regulatory region of human factor IX gene. J Biol Chem. 1990 Apr 25;265(12):7062–7068. [PubMed] [Google Scholar]
  17. Sladek F. M., Zhong W. M., Lai E., Darnell J. E., Jr Liver-enriched transcription factor HNF-4 is a novel member of the steroid hormone receptor superfamily. Genes Dev. 1990 Dec;4(12B):2353–2365. doi: 10.1101/gad.4.12b.2353. [DOI] [PubMed] [Google Scholar]
  18. Thompson A. R. Structure, function, and molecular defects of factor IX. Blood. 1986 Mar;67(3):565–572. [PubMed] [Google Scholar]
  19. Wang L. H., Tsai S. Y., Sagami I., Tsai M. J., O'Malley B. W. Purification and characterization of chicken ovalbumin upstream promoter transcription factor from HeLa cells. J Biol Chem. 1987 Nov 25;262(33):16080–16086. [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