Induction of apoptosis is driven by nuclear retention of protein kinase C delta.
Protein kinase C delta (PKC delta) mediates apoptosis downstream of many apoptotic stimuli. Because of its ubiquitous expression, tight regulation of the proapoptotic function of PKC delta is critical for cell survival. Full-length PKC delta is found in all cells, whereas the catalytic fragment of PKC delta, generated by caspase cleavage, is only present in cells undergoing apoptosis. Here we show that full-length PKC delta transiently accumulates in the nucleus in response to etoposide and that nuclear translocation precedes caspase cleavage of PKC delta. Nuclear PKC delta is either cleaved by caspase 3, resulting in accumulation of the catalytic fragment in the nucleus, or rapidly exported by a Crm1-sensitive pathway, thereby assuring that sustained nuclear accumulation of PKC delta is coupled to caspase activation. Nuclear accumulation of PKC delta is necessary for caspase cleavage, as mutants of PKC delta that do not translocate to the nucleus are not cleaved. However, caspase cleavage of PKC delta per se is not required for apoptosis, as an uncleavable form of PKC delta induces apoptosis when retained in the nucleus by the addition of an SV-40 nuclear localization signal. Finally, we show that kinase negative full-length PKC delta does not translocate to the nucleus in apoptotic cells but instead inhibits apoptosis by blocking nuclear import of endogenous PKC delta. These studies demonstrate that generation of the PKC delta catalytic fragment is a critical step for commitment to apoptosis and that nuclear import and export of PKC delta plays a key role in regulating the survival/death pathway.
DeVries-Seimon
Tracie A
TA
Department of Medicine, Columbia University, New York, NY 10032, USA.
Ohm
Angela M
AM
Humphries
Michael J
MJ
Reyland
Mary E
ME
D021581:Active Transport, Cell Nucleus; D017209:Apoptosis; D002384:Catalysis; D002467:Cell Nucleus / Q000378:metabolism*; D002470:Cell Survival; D053938:DNA Fragmentation; D004789:Enzyme Activation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D051745:Protein Kinase C-delta / Q000378:metabolism*; D021381:Protein Transport; D013997:Time Factors
D049452:Green Fluorescent Proteins; D051745:Protein Kinase C-delta