Inducible transcription factor expression in a cell culture model of apoptosis.
We have developed a model of nerve cell death based on the toxicity of okadaic acid, a compound that triggers apoptosis in PC12 cells via a protein synthesis-dependent mechanism. The cell death process is accompanied by induction of JunB, c-Jun, JunD and Fos proteins. Phosphorylation-specific antibodies were used to demonstrate that c-Jun is phosphorylated at serine 63 and serine 73. Electrophoretic gel mobility shift and pAP1-Luc luciferase assays showed that expression of ITFs is associated with increases in AP-1 binding and in AP-1 transcriptional activity. In addition, dose response and time course studies provided strong correlative evidence that Fos and Jun proteins are involved in the apoptotic death cascades. Thus, this model provides a useful system to investigate the role of inducible transcription factor proteins in apoptosis.
Woodgate
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The Department of Pharmacology and Clinical Pharmacology, Faculty of Medicine and Health Science, University of Auckland, Private Bag 92019, Auckland, New Zealand.
Walton
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MacGibbon
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GA
Dragunow
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D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D015342:DNA Probes; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*; D019319:Okadaic Acid / Q000494:pharmacology; D016716:PC12 Cells; D000073718:Pancreatitis-Associated Proteins; D010766:Phosphorylation; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D051381:Rats; D018808:Transcription Factor AP-1 / Q000235:genetics* / Q000378:metabolism
D017362:Cyclic AMP Response Element-Binding Protein; D015342:DNA Probes; D004791:Enzyme Inhibitors; D000073718:Pancreatitis-Associated Proteins; D016755:Proto-Oncogene Proteins c-jun; C000612141:REG3A protein, human; D018808:Transcription Factor AP-1; D019319:Okadaic Acid