Direct inhibition of c-Jun N-terminal kinase in sympathetic neurones prevents c-jun promoter activation and NGF withdrawal-induced death

J Neurochem. 2001 Mar;76(5):1439-54. doi: 10.1046/j.1471-4159.2001.00150.x.

Abstract

c-Jun N-terminal kinases (JNKs) regulate gene expression by phosphorylating transcription factors, such as c-Jun. Studies with JNK: knockout mice suggest that JNK activity may be required for excitotoxin-induced apoptosis in the adult hippocampus and for apoptosis in the developing embryonic neural tube. Here we investigate the role of JNKs in classical neurotrophin-regulated developmental neuronal death by using nerve growth factor (NGF)-dependent sympathetic neurones. In this system, NGF withdrawal leads to an increase in JNK activity, an increase in c-Jun protein levels and c-Jun N-terminal phosphorylation before the cell death commitment point, and c-Jun activity is required for cell death. To inhibit JNK activity in sympathetic neurones we have used two different JNK inhibitors that act by distinct mechanisms: the compound SB 203580 and the JNK binding domain (JBD) of JNK interacting protein 1 (JIP-1). We demonstrate that JNK activity is required for c-Jun phosphorylation, c-jun promoter activation and NGF withdrawal-induced apoptosis. We also show that ATF-2, a c-Jun dimerization partner that can regulate c-jun gene expression, is activated following NGF deprivation. Finally, by co-expressing the JBD and a regulatable c-Jun dominant negative mutant we demonstrate that JNK and AP-1 function in the same pro-apoptotic signalling pathway after NGF withdrawal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cell Survival / drug effects
  • Cells, Cultured
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation, Enzymologic*
  • Genes, jun*
  • Imidazoles / pharmacology
  • JNK Mitogen-Activated Protein Kinases
  • Mice
  • Mice, Knockout
  • Mitogen-Activated Protein Kinase 10
  • Mitogen-Activated Protein Kinase 8
  • Mitogen-Activated Protein Kinase 9
  • Mitogen-Activated Protein Kinases / genetics*
  • Mitogen-Activated Protein Kinases / metabolism*
  • Nerve Growth Factor / pharmacology*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / physiology*
  • Promoter Regions, Genetic*
  • Protein-Tyrosine Kinases / genetics
  • Proto-Oncogene Proteins c-jun / genetics
  • Pyridines / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction
  • Superior Cervical Ganglion / cytology
  • Superior Cervical Ganglion / physiology*
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • Enzyme Inhibitors
  • Imidazoles
  • Proto-Oncogene Proteins c-jun
  • Pyridines
  • Transcription Factors
  • Nerve Growth Factor
  • Mitogen-Activated Protein Kinase 10
  • Mitogen-Activated Protein Kinase 9
  • Protein-Tyrosine Kinases
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinase 8
  • Mitogen-Activated Protein Kinases
  • SB 203580