PKCε promotes HuD-mediated neprilysin mRNA stability and enhances neprilysin-induced Aβ degradation in brain neurons.
Amyloid-beta (Aβ) peptide accumulation in the brain is a pathological hallmark of all forms of Alzheimer's disease. An imbalance between Aβ production and clearance from the brain may contribute to accumulation of neurotoxic Aβ and subsequent synaptic loss, which is the strongest correlate of the extent of memory loss in AD. The activity of neprilysin (NEP), a potent Aβ-degrading enzyme, is decreased in the AD brain. Expression of HuD, an mRNA-binding protein important for synaptogenesis and neuronal plasticity, is also decreased in the AD brain. HuD is regulated by protein kinase Cε (PKCε), and we previously demonstrated that PKCε activation decreases Aβ levels. We hypothesized that PKCε acts through HuD to stabilize NEP mRNA, modulate its localization, and support NEP activity. Conversely, loss of PKCε-activated HuD in AD leads to decreased NEP activity and accumulation of Aβ. Here we show that HuD is associated with NEP mRNA in cultures of human SK-N-SH cells. Treatment with bryostatin, a PKCε-selective activator, enhanced NEP association with HuD and increased NEP mRNA stability. Activation of PKCε also increased NEP protein levels, increased NEP phosphorylation, and induced cell surface expression. In addition, specific PKCε activation directly stimulated NEP activity, leading to degradation of a monomeric form of Aβ peptide and decreased Aβ neuronal toxicity, as measured by cell viability. Bryostatin treatment also rescued Aβ-mediated inhibition of HuD-NEP mRNA binding, NEP protein expression, and NEP cell membrane translocation. These results suggest that PKCε activation reduces Aβ by up-regulating, via the mRNA-binding protein HuD, Aβ-degrading enzymes such as NEP. Thus, PKCε activation may have therapeutic efficacy for AD by reducing neurotoxic Aβ accumulation as well as having direct anti-apoptotic and synaptogenic effects.
Blanchette Rockefeller Neurosciences Institute at West Virginia University, Morgantown, West Virginia, United States of America.
Chol Seung
CS
Lim
Blanchette Rockefeller Neurosciences Institute at West Virginia University, Morgantown, West Virginia, United States of America.
Daniel L
DL
Alkon
D016229:Amyloid beta-Peptides / Q000378:metabolism* / Q000633:toxicity; D001921:Brain / Q000166:cytology*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D051959:ELAV Proteins / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D015260:Neprilysin / Q000235:genetics*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D051744:Protein Kinase C-epsilon / Q000378:metabolism*; D021381:Protein Transport; D059748:Proteolysis; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics
D016229:Amyloid beta-Peptides; D051959:ELAV Proteins; D012333:RNA, Messenger; D051744:Protein Kinase C-epsilon; D015260:Neprilysin