DNA synthesis and neuronal apoptosis caused by familial Alzheimer disease mutants of the amyloid precursor protein are mediated by the p21 activated kinase PAK3.
Apoptotic pathways and DNA synthesis are activated in neurons in the brains of individuals with Alzheimer disease (AD). However, the signaling mechanisms that mediate these events have not been defined. We show that expression of familial AD (FAD) mutants of the amyloid precursor protein (APP) in primary neurons in culture causes apoptosis and DNA synthesis. Both the apoptosis and the DNA synthesis are mediated by the p21 activated kinase PAK3, a serine-threonine kinase that interacts with APP. A dominant-negative kinase mutant of PAK3 inhibits the neuronal apoptosis and DNA synthesis; this effect is abolished by deletion of the PAK3 APP-binding domain or by coexpression of a peptide representing this binding domain. The involvement of PAK3 specifically in FAD APP-mediated apoptosis rather than in general apoptotic pathways is suggested by the facts that a dominant-positive mutant of PAK3 does not alone cause neuronal apoptosis and that the dominant-negative mutant of PAK3 does not inhibit chemically induced apoptosis. Pertussis toxin, which inactivates the heterotrimeric G-proteins Go and Gi, inhibits the apoptosis and DNA synthesis caused by FAD APP mutants; the apoptosis and DNA synthesis are rescued by coexpression of a pertussis toxin-insensitive Go. FAD APP-mediated DNA synthesis precedes FAD APP-mediated apoptosis in neurons, and inhibition of neuronal entry into the cell cycle inhibits the apoptosis. These data suggest that a normal signaling pathway mediated by the interaction of APP, PAK3, and Go is constitutively activated in neurons by FAD mutations in APP and that this activation causes cell cycle entry and consequent apoptosis.
McPhie
Donna L
DL
Department of Psychiatry, Harvard Medical School and McLean Hospital, Belmont, Massachusetts 02478, USA. mcphie@helix.mgh.harvard.edu
Coopersmith
Robert
R
Hines-Peralta
Andrew
A
Chen
Yuzhi
Y
Ivins
Kathryn J
KJ
Manly
Susan P
SP
Kozlowski
Michael R
MR
Neve
Kim A
KA
Neve
Rachael L
RL
D000544:Alzheimer Disease / Q000235:genetics*; D016564:Amyloid beta-Protein Precursor / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001973:Bromodeoxyuridine; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis*; D005799:Genes, Dominant; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D009154:Mutation; D009420:Nervous System / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009928:Organ Specificity; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology; D054462:p21-Activated Kinases; D020691:rab GTP-Binding Proteins / Q000096:biosynthesis; D020696:rab5 GTP-Binding Proteins / Q000096:biosynthesis; D000099193:rab11 GTP-Binding Proteins
D016564:Amyloid beta-Protein Precursor; D001973:Bromodeoxyuridine; D004247:DNA; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D054462:p21-Activated Kinases; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C515913:Pak3 protein, rat