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.

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.

• Authors: A G Smith, G Brightwell, S E Smit, P G Parsons, R A Sturm
• Journal: Melanoma research (Melanoma Res) Vol. 8 Issue 1 Pg. 2-10 (Feb 1998) ISSN: 0960-8931 [Print] England
• PMID: 9508370 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

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

Topics

• Animals
• Blotting, Western
• COS Cells
• Cell Nucleus (chemistry, genetics)
• DNA, Neoplasm (isolation & purification, 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