Tau is phosphorylated by GSK-3 at several sites found in Alzheimer disease and its biological activity markedly inhibited only after it is prephosphorylated by A-kinase.
Alzheimer disease is characterized by a specific type of neuronal degeneration in which the microtubule associated protein tau is abnormally hyperphosphorylated causing the disruption of the microtubule network. We have found that the phosphorylation of human tau (tau3L) by A-kinase, GSK-3 or CK-1 inhibits its microtubule assembly-promoting and microtubule-binding activities. However, the inhibition of these activities of tau by GSK-3 is significantly increased if tau is prephosphorylated by A-kinase or CK-1. The most potent inhibition is observed by combination phosphorylation of tau with A-kinase and GSK-3. Under these conditions, only very few microtubules are seen by electron microscopy. Sequencing of 32P-labeled trypsin phosphopeptides from tau prephosphorylated by A-kinase (using unlabeled ATP) and further phosphorylated by GSK-3 in the presence of [gamma-32P]ATP revealed that Ser-195, Ser-198, Ser-199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-356 and Ser-404 are phosphorylated, whereas if tau is not prephosphorylated by A-kinase, GSK-3 phosphorylates it at Thr-181, Ser-184, Ser-262, Ser-356 and Ser-400. These data suggest that (i) prephosphorylation of tau by A-kinase makes additional and different sites accessible for phosphorylation by GSK-3; (ii) phosphorylation of tau at these additional sites further inhibits the biological activity of tau in its ability to bind to microtubules and promote microtubule assembly. Thus a combined role of A-kinase and GSK-3 should be considered in Alzheimer neurofibrillary degeneration.
Chemical Neuropathology Department, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314-6399, USA.
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Smith
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D000544:Alzheimer Disease / Q000378:metabolism*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D047388:Casein Kinases; D002853:Chromatography, Liquid / Q000379:methods; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3; D006801:Humans; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism*
D000596:Amino Acids; D010761:Phosphorus Radioisotopes; D016875:tau Proteins; D011494:Protein Kinases; D047388:Casein Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D038362:Glycogen Synthase Kinase 3