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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #183882
Title: PROTEOLYTIC ACTIVATION OF PKC-DELTA CONTRIBUTES TO OXIDATIVE- AND ER-STRESS INDUCED APOPTOTIC CELL DEATH IN PRP*SEN-OVEREXPRESSING MOUSE NEURONAL CELLS

Author
item ANANTHARAM, VELLAREDDY - IOWA STATE UNIVERSITY
item VORBERG, INA - IOWA STATE UNIVERSITY
item CHOI, CHRIS - UNIVERSITY OF MUNICH
item KANTHASAMY, ARTHI - IOWA STATE UNIVERSITY
item Richt, Juergen
item PRIOLA, SUZETTE - ROCKY MOUNTAIN LABS, NIH
item KANTHASAMY, ANUMANTHA - IOWA STATE UNIVERSITY

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/29/2004
Publication Date: 10/14/2004
Citation: Anantharam, V., Vorberg, I., Choi, C., Kanthasamy, A., Richt, J.A., Priola, S.A., Kanthasamy, A.G. 2004. Proteolytic activation of PKC-delta contributes to oxidative- and ER-stress induced apoptotic cell death in PrP*sen-overexpressing mouse neuronal cells [abstract]. Animal Prion Diseases and the Americas. p. 48.

Interpretive Summary:

Technical Abstract: Although prion protein is abundantly expressed in the CNS, its biological function has not yet been established. To determine the role of prion protein in oxidative stress and protein processing, we compared the effects of H*2O*2 and the endoplasmic reticulum (ER) stress-inducers brefeldin A and tunicamycin on apoptotic cell signaling in a neural cell line derived from PrP knockout mice engineered to stably express non-pathogenic mouse prion protein (PrP*sen) and empty vector-expressing prion knockout cells (PrP*ko). Treatment with H*2O*2 (10-100 uM), brefeldin A (3-30 uM), and tunicamycin (0.6-6 uM) resulted in dose- and time-dependent increases in cytotoxic cell death, cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation over a 24 hr period in both PrP*sen and PrP*ko cells. However, ER stress-induced caspase-3 activation, DNA fragmentation, and cytotoxic cell death were significantly higher in PrP*sen than PrP*ko. Alternatively, H*2O*2 stress-induced caspase-3 activation, DNA fragmentation, and cytotoxic cell death were significantly reduced in PrP*sen as compared to PrP*ko, suggesting that prion protein may be neuroprotective against oxidative damage. Pretreatment with 2 uM MnTBAP (SOD mimetic, ROS inhibitor) suppressed H*2O*2 stress but not ER stress-induced apoptotic cell death. Additionally, caspase-12 and -8 were activated only in cells treated with brefeldin A and tunicamycin but not with H*2O*2. Interestingly, PKC-delta, an oxidative stress sensitive kinase, was proteolytically cleaved and activated by caspase-3 during both oxidative stress and ER stress. Comparative analysis of PKC-delta proteolytic activation in PrP*sen and PrP*ko revealed that prion expressing cells were protected from oxidative stress-induced PKC-delta activation; however, exacerbated ER stress induced PKC-delta activation. Overexpression of the kinase inactive PKC-delta dominant negative mutant (PKC-delta**K376R) or the caspase-3 cleavage site-resistant PKC-delta mutant (PKC-delta**D327A) significantly attenuated both ER- and oxidative stress-induced apoptotic cell death. Together, these results suggest that prion protein enhances the susceptibility of neural cells to impairment in protein processing and trafficking but decreases the vulnerability to oxidative insults, and that PKC-delta is a key downstream mediator of cellular stress-induced neuronal apoptosis.