Elsevier

Brain Research

Volume 611, Issue 2, 21 May 1993, Pages 237-242
Brain Research

Phosphorylation of tau by proline-directed protein kinase (p34cdc2/p58cyclin A) decreases tau-induced microtubule assembly and antibody SMI33 reactivity

https://doi.org/10.1016/0006-8993(93)90508-KGet rights and content

Abstract

Tau protein was evaluated as a substrate for a proline-directed protein kinase (p34cdc2/p58cyclin A) which recognizes the phosphorylation site motif X-Ser/Thr-Pro-X. The shortest human tau isoform, expressed as a recombinant protein, was phosphorylated to a stoichiometry of 2 mol phosphate/mol tau. Phosphoamino acid analysis revealed phosphorylation of both serine and threonine residues. Phosphorylation of recombinant tau resulted in a decreased ability to induce microtubule assembly but had no effect on the final extent of microtubule formation or on the rate of cold-induced microtubule disassembly. Phosphorylation of tau by the proline-directed protein kinase completely blocked immunoreactivity with antibody SMI33. Phosphorylation did not create the epitopes for the phosphate-dependent antibodies SMI31 or SMI34. Antibody SMI33 recognizes neurofibrillary tangles after treatment with alkaline phosphatase, suggesting that the proline-directed protein kinase may phosphorylate tau at sites that are phosphorylated in Alzheimer's disease.

References (42)

  • HallF.L. et al.

    Proline-directed protein phosphorylation and cell cycle regulation

    Curr. Opinion Cell Biol.

    (1991)
  • HoshiM. et al.

    Protein kinase C phosphorylates tau and induces its functional alterations

    FEBS Lett.

    (1987)
  • MurphyD.B.

    Assembly-disassembly purification and characterization of microtubule protein without glycerol

    Methods Cell Biol.

    (1982)
  • ReimannE.M. et al.

    Purification and properties of rabbit skeletal muscle adenosine 3′,5′0monophosphate dependent protein kinases

    J. Biol. Chem.

    (1971)
  • ScottC.W. et al.

    Phosphorylation of recombinant tau by cAMP-dependent protein kinase: identification of phosphorylation sites and effect on microtubule assembly

    J. Biol. Chem.

    (1993)
  • VullietP.R. et al.

    Identification of a novel proline-directed serine/threonine protein kinase in rat pheochromocytoma

    J. Biol. Chem.

    (1989)
  • AhnN.G. et al.

    Identification of an activator of the microtubule-associated protein kinases ERKI and ERK2 in PC12 cells stimulated with nerve growth factor or bradykinin

    J. Neurochem.

    (1992)
  • BiernatJ. et al.

    The switch of tau-protein to an Alzheimer-like state includes the phosphorylation of two serine-proline motifs upstream of the microtubule binding region

    EMBO J.

    (1992)
  • BraunR.K. et al.

    Tissue distribution of a novel growth factor sensitive protein kinase

  • ColemanM.P. et al.

    Phosphate-dependent monoclonal antibodies to neurofilaments and Alzheimer neurofibrillary tangles recognize a synthetic phosphopeptide

    J. Neurochem.

    (1990)
  • CorkL.C. et al.

    Phosphorylated neurofilament antigens in neurofibrillary tangles in Alzheimer's disease

    J. Neuropathol. Exp. Neurol.

    (1986)
  • Cited by (37)

    • Tau pathophysiology in neurodegeneration: a tangled issue

      2009, Trends in Neurosciences
      Citation Excerpt :

      With tau containing >80 possible phosphorylation sites in its primary amino acid structure, nearly 20% of the molecule can be phosphorylated and a large number of protein kinases (>20) and phosphatases have been identified that control tau phosphorylation [66]. Some of the most widely studied kinases and phosphatases that have been implicated in the pathological phosphorylation of tau are glycogen synthase kinase 3β (GSK3β) [67,68], microtubule affinity regulating kinase (MARK) [69], mitogen activated protein kinase [70] and cyclin dependent kinase 5 (Cdk5) [71–75], with protein phosphatase 2A being the most important tau phosphatase [76–78]. With so many phosphorylation sites and kinases and phosphatases regulating the phosphorylation state of tau, it is clear that the phosphorylation state of tau is very highly regulated and important in its normal and pathological function.

    View all citing articles on Scopus
    View full text