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.
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.
Smith
A G
AG
Centre for Molecular and Cellular Biology, University of Queensland, Brisbane, Australia.
Brightwell
G
G
Smit
S E
SE
Parsons
P G
PG
Sturm
R A
RA
D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics; D004273:DNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003958:Diamide / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006367:HeLa Cells; D018398:Homeodomain Proteins; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008545:Melanoma / Q000235:genetics*; D008546:Melanoma, Experimental / Q000235:genetics*; D008670:Metals; D051379:Mice; D010084:Oxidation-Reduction; D050809:POU Domain Factors; D011817:Rabbits; D014157:Transcription Factors / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D008670:Metals; D050809:POU Domain Factors; D014157:Transcription Factors; C085076:transcription factor Brn-2; D003958:Diamide; D006861:Hydrogen Peroxide