Akt/PKB kinase phosphorylates separately Thr212 and Ser214 of tau protein in vitro.
Microtubule-associated protein tau contains a consensus motif for protein kinase B/Akt (Akt), which plays an essential role in anti-apoptotic signaling. The motif encompasses the AT100 double phospho-epitope (Thr212/Ser214), a specific marker for Alzheimer's disease (AD) and other neurodegenerations, raising the possibility that it could be generated by Akt. We studied Akt-dependent phosphorylation of tau protein in vitro. We found that Akt phosphorylated both Thr212 and Ser214 in the longest and shortest tau isoforms as determined using phospho site-specific antibodies against tau. Akt did not phosphorylate other tau epitopes, including Tau-1, AT8, AT180, 12E8 and PHF-1. The Akt-phosphorylated tau retained its initial electrophoretic mobility. Immunoprecipitation studies with phospho-specific Thr212 and Ser214 antibodies revealed that only one of the two sites is phosphorylated per single tau molecule, resulting in tau immunonegative for AT100. Mixed kinase studies showed that prior Ser214 phosphorylation by Akt blocked protein kinase A but not GSK3beta activity. On the other hand, GSK3beta selectively blocked Ser214 phosphorylation, which was prevented by lithium. The results suggest that Akt may be involved in AD-specific phosphorylation of tau at the AT100 epitope in conjunction with other kinases. Our data suggest that phosphorylation of tau by Akt may play specific role(s) in Akt-mediated anti-apoptotic signaling, particularly relevant to AD and other neurodegenerations.
Ksiezak-Reding
Hanna
H
Neuroinflammation Research Laboratories, Department of Psychiatry, Mount Sinai School of Medicine, Box 1230, One Gustave L. Levy Place, New York, NY 10029, USA. hanna.reding@mssm.edu
Pyo
Han Kyoung
HK
Feinstein
Boris
B
Pasinetti
Giulio M
GM
D000544:Alzheimer Disease / Q000175:diagnosis; D000595:Amino Acid Sequence; D015415:Biomarkers; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D007700:Kinetics; D009410:Nerve Degeneration / Q000175:diagnosis; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D010769:Phosphothreonine / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
D015415:Biomarkers; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D016875:tau Proteins; D010769:Phosphothreonine; D010768:Phosphoserine; C494918:AKT1 protein, human; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3