Molecular and Cellular Biochemistry

, Volume 229, Issue 1, pp 73–83

The characterization and purification of a human transcription factor modulating the glutathione peroxidase gene in response to oxygen tension

  • Frank Merante
  • Svetlana M. Altamentova
  • Donald A.G. Mickle
  • Richard D. Weisel
  • Bradley J. Thatcher
  • Brian M. Martin
  • John G. Marshall
  • Laura C. Tumiati
  • Douglas B. Cowan
  • Ren-Ke Li
Article

DOI: 10.1023/A:1017921110363

Cite this article as:
Merante, F., Altamentova, S.M., Mickle, D.A. et al. Mol Cell Biochem (2002) 229: 73. doi:10.1023/A:1017921110363

Abstract

An oxygen responsive transcription factor regulating human glutathione peroxidase gene (GPx) through two oxygen responsive elements (ORE1 and ORE2) has been purified and characterized by sequence-specific DNA affinity chromatography. The DNA binding activity, termed Oxygen Responsive Element Binding Protein (OREBP), was partially represented by a 77 kD polypeptide (p70) possessing a blocked N-terminus. The p70 subunit co-eluted with an 86 kD subunit (p80) from affinity columns. N-terminal sequencing analysis of the 86 kD component revealed that this protein represented the larger member of the Ku antigen complex. The identity of the purified 77 kD subunit was determined by Western blot analysis using an antibody directed against the p70 protein. In addition to binding the GPx-ORE, the OREBP was itself regulated by oxygen tension. It was found that the abundance of the ORE binding activity was decreased in cells maintained at low oxygen tension (40 mm Hg). Anti-Ku-antibodies specifically supershifted the OREBP-ORE DNA complex. These observations further add to the numerous nuclear roles of the Ku-transcription factor.

DNA binding proteinsglutathione peroxidasehypoxiaKu-antigenprotein purificationtranscription factors

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Frank Merante
    • 1
  • Svetlana M. Altamentova
    • 2
  • Donald A.G. Mickle
    • 2
  • Richard D. Weisel
    • 2
  • Bradley J. Thatcher
    • 3
  • Brian M. Martin
    • 3
  • John G. Marshall
    • 3
  • Laura C. Tumiati
    • 2
  • Douglas B. Cowan
    • 2
  • Ren-Ke Li
    • 2
  1. 1.Tm Bioscience CorporationToronto, OntarioCanada
  2. 2.Centre for Cardiovascular Research, Department of Clinical Biochemistry and the Division of Cardiovascular Surgery, The Toronto HospitalUniversity of TorontoToronto, OntarioCanada
  3. 3.BethesdaNational Institutes of Medical Health (NIH)USA