Caspase-dependent, geldanamycin-enhanced cleavage of co-chaperone p23 in leukemic apoptosis.
Co-chaperone p23 is a component of the heat-shock protein (Hsp)90 multiprotein-complex and is an important modulator of Hsp90 activity. Hsp90 client proteins involved in oncogenic survival signaling are frequently mutated in leukemia, and the integrity of the Hsp90 complex could therefore be important for leukemic cell survival. We demonstrate here that p23 is cleaved to a stable 17 kDa fragment in leukemic cell lines treated with commonly used chemotherapeutic drugs. The cleavage of p23 paralleled the activation of procaspase-7 and -3 and was suppressed by the caspase-3/-7 inhibitor DEVD-FMK. In vitro translated 35S-p23 (in reticulocyte lysate) was cleaved at D142 and D145 by caspase-7 and -3. Cleavage of p23 occurred in caspase-3-deficient MCF-7 cells, suggesting a role for caspase-7 in intact cells. The Hsp90 inhibitor geldanamycin enhanced caspase-dependent p23 cleavage both in vitro and in intact cells. Geldanamycin also enhanced anthracycline-induced caspase activation and apoptosis. We conclude that p23 is a prominent target in leukemic cell apoptosis. Geldanamycin enhanced p23 cleavage both by rendering p23 more susceptible to caspases and by enhancing chemotherapy-induced caspase activation. These findings underscore the importance of the Hsp90-complex in antileukemic treatment, and suggest that p23 may have a role in survival signaling.
Gausdal
G
G
Department of Biomedicine, Section of Anatomy and Cell Biology and PROBE, University of Bergen, Norway.
Gjertsen
B T
BT
Fladmark
K E
KE
Demol
H
H
Vandekerckhove
J
J
Døskeland
S-O
SO
D017209:Apoptosis; D016227:Benzoquinones; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D047029:Lactams, Macrocyclic; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology; D018832:Molecular Chaperones / Q000187:drug effects / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D010750:Phosphoproteins / Q000187:drug effects / Q000378:metabolism*; D000071876:Prostaglandin-E Synthases; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011809:Quinones / Q000494:pharmacology*; D011980:Receptors, Progesterone / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism
D016227:Benzoquinones; D004791:Enzyme Inhibitors; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D010750:Phosphoproteins; D011809:Quinones; D011980:Receptors, Progesterone; D011505:Protein-Tyrosine Kinases; C505407:CASP3 protein, human; C505520:CASP7 protein, human; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases; C490849:PTGES3 protein, human; D000071876:Prostaglandin-E Synthases; C001277:geldanamycin