Abstract
The sIgG(+) lymphoblastoid B cell line CESS spontaneously produces a high amount of NGF and expresses both high affinity (p140(Trk-A)) and low affinity (p75(NTR)) NGF receptors. Blocking NGF signals with neutralizing antibodies or specific Trk-A inhibitors induces a rapid phosphorylation of antiapoptotic Bcl-2 protein, followed by caspase activation, and apoptotic death of CESS cells. Bcl-2 phosphorylation in several sites within a approximately 60 aa "loop" domain of protein is known to regulate its antiapoptotic function. Accordingly, CESS cells expressing the loop deletional mutant cDNA constructs Bcl-2 Delta40-91 were completely resistant to apoptosis induced by NGF withdrawal, indicating that Bcl-2 phosphorylation is a critical event. NGF withdrawal induces p38 MAPK, but not JNK, activation in CESS cells, and SB203580, a specific inhibitor of p38 MAPK, is able to prevent both Bcl-2 phosphorylation and apoptosis, indicating that p38 MAPK is the enzyme responsible for these events.
Copyright 2000 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Apoptosis / drug effects*
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Apoptosis / physiology
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B-Lymphocytes
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Carbazoles / pharmacology
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Cell Division / drug effects
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Enzyme Activation
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Enzyme Inhibitors / pharmacology
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Genes, bcl-2
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Humans
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Indole Alkaloids
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JNK Mitogen-Activated Protein Kinases
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Kinetics
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Mitogen-Activated Protein Kinases / metabolism*
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Nerve Growth Factor / pharmacology*
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Phosphorylation
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Proto-Oncogene Proteins c-bcl-2 / metabolism*
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Receptor, trkA / genetics
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Receptor, trkA / physiology
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Receptors, Nerve Growth Factor / genetics
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Receptors, Nerve Growth Factor / physiology
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Sequence Deletion
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Signal Transduction / drug effects
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Signal Transduction / physiology
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Tumor Cells, Cultured
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Tyrphostins / pharmacology
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p38 Mitogen-Activated Protein Kinases
Substances
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AG-879
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Carbazoles
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Enzyme Inhibitors
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Indole Alkaloids
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Proto-Oncogene Proteins c-bcl-2
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Receptors, Nerve Growth Factor
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Tyrphostins
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Nerve Growth Factor
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staurosporine aglycone
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Receptor, trkA
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JNK Mitogen-Activated Protein Kinases
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Mitogen-Activated Protein Kinases
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p38 Mitogen-Activated Protein Kinases