A novel mechanism of non-Aβ component of Alzheimer's disease amyloid (NAC) neurotoxicity. Interplay between p53 protein and cyclin-dependent kinase 5 (Cdk5).
The non-Aβ component of Alzheimer's disease (AD) amyloid (NAC) is produced from the precursor protein NACP/α-synuclein (ASN) by till now unknown mechanism. Previous study showed that like ASN, NAC peptide induced oxidative/nitrosative stress and apoptosis. Our present study focused on the mechanisms of PC12 cells death evoked by NAC peptide, with particular consideration on the role of p53 protein. On the basis of molecular and transmission electron microscopic (TEM) analysis it was found that exogenous NAC peptide (10 μM) caused mitochondria dysfunction, enhanced free radical generation, and induced both apoptotic and autophagic cell death. Morphological and immunocytochemical evidence from TEM showed marked changes in expression and in translocation of proapoptotic protein Bax. We also observed time-dependent enhancement of Tp53 gene expression after NAC treatment. Free radicals scavenger N-tert-butyl-alpha-phenylnitrone (PBN, 1 mM) and p53 inhibitor (α-Pifithrin, 20 μM) significantly protected PC12 cells against NAC peptide-evoked cell death. In addition, exposure to NAC peptide resulted in higher expression of cyclin-dependent kinase 5 (Cdk5), one of the enzymes responsible for p53 phosphorylation and activation. Concomitantly, we observed the increase of expression of Cdk5r1 and Cdk5r2 genes, coding p35 and p39 peptides that are essential regulators of Cdk5 activity. Moreover, the specific Cdk5 inhibitor (BML-259, 10 μM) protected large population of cells against NAC-evoked cell death. Our findings indicate that NAC peptide exerts its toxic effect by activation of p53/Cdk5 and Bax-dependent apoptotic signaling pathway.
Mossakowski Medical Research Center, Polish Academy of Sciences, Department of Cellular Signaling, Pawińskiego 5, 02-106 Warsaw, Poland. aniakazmierczak@gmail.com
Anna
A
Kaźmierczak
Grzegorz A
GA
Czapski
Agata
A
Adamczyk
Barbara
B
Gajkowska
Joanna B
JB
Strosznajder
D000544:Alzheimer Disease / Q000378:metabolism* / Q000503:physiopathology; D000682:Amyloid / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D051360:Cyclin-Dependent Kinase 5 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D016716:PC12 Cells; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D051844:alpha-Synuclein / Q000378:metabolism* / Q000633:toxicity; D051028:bcl-2-Associated X Protein / Q000187:drug effects / Q000378:metabolism
D000682:Amyloid; D016166:Free Radical Scavengers; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D051844:alpha-Synuclein; D051028:bcl-2-Associated X Protein; D051360:Cyclin-Dependent Kinase 5; C495907:Cdk5 protein, rat