Site-specific dephosphorylation of tau protein at Ser202/Thr205 in response to microtubule depolymerization in cultured human neurons involves protein phosphatase 2A.
Tau proteins isolated from paired helical filaments, the major building blocks of Alzheimer's disease neurofibrillary tangle, are abnormally phosphorylated and unable to bind microtubules. To examine the dynamics of tau phosphorylation and to identify specific tau phosphorylation sites involved in the stabilization of microtubules, we treated cultured postmitotic neuron-like cells (NT2N) derived from a human teratocarcinoma cell line (NTera2/D1) with drugs that depolymerize microtubules (i.e. colchicine or nocodazole). This led to the recovery of dephosphorylated tau from the NT2N cells as monitored by a relative increase in the electrophoretic mobility of tau and an increase in the turnover of [32P]PO4-labeled tau. However, not all phosphorylation sites on tau are affected by colchicine or nocodazole. Ser202/Thr205 appears to be completely and specifically dephosphorylated by protein phosphatase 2A since this dephosphorylation was blocked by inhibitors of protein phosphatase 2A but not by inhibitors of protein phosphatase 2B. These findings, together with the recent observation that protein phosphatase 2A is normally bound to microtubules in intact cells, suggest that the polymerization state of microtubules could modulate the phosphorylation state of tau at specific sites in the normal and Alzheimer's disease brain.
Merrick
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David Mahoney Institute of Neurological Sciences and the Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, 19104-4283, USA.
Demoise
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Lee
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D000544:Alzheimer Disease / Q000378:metabolism; D000595:Amino Acid Sequence; D000906:Antibodies; D001345:Autoradiography; D015153:Blotting, Western; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006801:Humans; D008715:Methionine / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D054648:Protein Phosphatase 2; D012694:Serine; D013462:Sulfur Radioisotopes; D018243:Teratocarcinoma; D013912:Threonine; D014407:Tumor Cells, Cultured; D016875:tau Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D000906:Antibodies; D000939:Epitopes; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D013462:Sulfur Radioisotopes; D016875:tau Proteins; D013912:Threonine; D012694:Serine; D008715:Methionine; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D015739:Nocodazole; D003078:Colchicine