Activator protein-1 DNA binding activation by hydrogen peroxide in neuronal and astrocytic primary cultures of trisomy-16 and diploid mice.
The effect of H(2)O(2) on DNA binding activity of activator protein-1 (AP-1) was studied by electrophoretic mobility shift assay (EMSA) in cortical primary cultures of trisomy-16 mice and their diploid littermates. Exposure to 10 microM H(2)O(2) for 15 min elicited a greater and earlier occurring increase of AP-1 DNA binding in neuronal primary cultures of trisomy-16 mice than of diploid mice. When astrocyte-rich primary cultures were exposed to 10 microM H(2)O(2) a two-fold increase of AP-1 DNA binding activity was found in trisomy-16 and diploid mice. Supershift EMSA analysis revealed that c-jun was a component of AP-1 in neuronal and glial cultures of diploid and trisomic mice. A 15-min exposure to 10 microM H(2)O(2) increased c-jun mRNA in cortical neuronal cultures by six-fold, compared with a two-fold increase in cultured astrocytes. The results documented that H(2)O(2)-elicited activation of AP-1 DNA binding in trisomy-16 primary cultures is transcriptionally regulated. Since oxidative stress also activates various stress-inducible protein kinases that may phosphorylate AP-1 dimers, the increase of AP-1 DNA binding may, in part, be triggered by phosphorylation.
Scortegagna
M
M
Laboratory of Molecular Immunology, NHLBI, National Institutes of Health, Bldg. 10, Room 7N312, Bethesda, MD 20892-1674, USA.
Galdzicki
Z
Z
Rapoport
S I
SI
Hanbauer
I
I
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004171:Diploidy; D004305:Dose-Response Relationship, Drug; D005260:Female; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D016755:Proto-Oncogene Proteins c-jun / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000187:drug effects* / Q000378:metabolism; D014314:Trisomy
D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; D004247:DNA; D006861:Hydrogen Peroxide