Redox regulation of Brn-2/N-Oct-3 POU domain DNA binding activity and proteolytic formation of N-Oct-5 during melanoma cell nuclear extraction

Melanoma Res. 1998 Feb;8(1):2-10. doi: 10.1097/00008390-199802000-00002.

Abstract

Reversible oxidation sensitivity of N-Oct-3 DNA binding activity was seen when melanoma extracts and recombinant Brn-2 protein were treated with a variety of metals, hydrogen peroxide and the cysteine disulphide bond forming agent diamide. Western blot analysis of diamide-oxidized N-Oct-3 protein indicated that this was likely to be due to intramolecular disulphide bonding. The potential role of oxidative loss of N-Oct-3 DNA binding activity is discussed in relation to redox changes that may occur during the early phase of apoptosis in neuronal cell lines and tissues. Brn-2 C-terminal antibody Western blot analysis of melanoma cell line nuclear extracts prepared using a combination of sodium dodecyl sulphate and NP-40 detergent cell lysis procedures demonstrated the formation of N-Oct-5 DNA binding activity via N-terminal proteolytic clipping of Brn-2/N-Oct-3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • COS Cells
  • Cell Nucleus / chemistry
  • Cell Nucleus / genetics
  • DNA, Neoplasm / isolation & purification
  • DNA, Neoplasm / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Diamide / pharmacology
  • Gene Expression Regulation, Neoplastic* / drug effects
  • HeLa Cells
  • Homeodomain Proteins
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Melanoma / genetics*
  • Melanoma, Experimental / genetics*
  • Metals
  • Mice
  • Oxidation-Reduction
  • POU Domain Factors
  • Rabbits
  • Transcription Factors / metabolism*
  • Tumor Cells, Cultured

Substances

  • DNA, Neoplasm
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • Metals
  • POU Domain Factors
  • Transcription Factors
  • transcription factor Brn-2
  • Diamide
  • Hydrogen Peroxide