Skip to main content

Non-proline-dependent protein kinases phosphorylate several sites found in tau from Alzheimer disease brain

Molecular and Cellular Biochemistry volume 154, pages 143–151 (1996)Cite this article

Abstract

Of 21 phosphorylation sites identified in PHF-tau 11 are on ser/thr-X motifs and are probably phosphorylated by non-proline-dependent protein kinases (non-PDPKs). The identities of the non-PDPKs and how they interact to hyperphosphorylate PHF-tau are still unclear. In a previous study we have shown that the rate of phosphorylation of human tau 39 by a PDPK (GSK-3) was increased several fold if tau were first prephosphorylated by non-PDPKs (Singh et al., FEBS Lett 358: 267-272, 1995). In this study we have examined how the specificity of a non-PDPK for different sites on human tau 39 is modulated when tau is prephosphorylated by other non-PDPKs (A-kinase, C-kinase, CK-1, CaM kinase II) as well as a PDPK (GSK-3). We found that the rate of phosphorylation of tau 39 by a non-PDPK can be stimulated if tau were first prephosphorylated by other non-PDPKs. Of the four non-PDPKs only CK-1 can phosphorylate sites (thr 231, ser 396, ser 404) known to be present in PHF-tau. Further, these sites were phosphorylated more rapidly and to a greater extent by CK-1 if tau 39 were first prephosphorylated by A-kinase, CaM kinase II or GSK-3. These results suggest that the site specificities of the non-PDPKs that participate in PHF-tau hyperphosphorylation can be modulated at the substrate level by the phosphorylation state of tau.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

39,95 €

Price includes VAT (Finland)

Abbreviations

PHF:

paired helical filaments

A-kinase:

cyclic AMP-dependent protein kinase

CaM kinase II:

calcium/calmodulin-dependent protein kinase II

C-kinase:

calcium/phospholipid-dependent protein kinase

CK-1:

casein kinase-1

CK-2:

casein kinase-2

GSK-3:

glycogen synthase kinase-3

MAP kinase:

mitogen-activated protein kinase

PDPK:

proline-dependent protein kinase

References

  1. Lindwall G, Cole RD: Phosphorylation affects the ability of tau protein to promote microtubule assembly. J Biol Chem 259: 5301–5305, 1984

    Google Scholar 

  2. Biernat J, Gustke N, Drewes G, Mandelkow EM, Mandelkow E: Phosphorylation of Ser262 strongly reduces binding of tau to microtubules: Distinction between PHF-like immunoreactivity and microtubule binding. Neuron 11: 153–163, 1993

    Google Scholar 

  3. Grundke-Iqbal I, Iqbal K, Quinlan M, Tung Y-C, Zaidi MS, Wisniewski HM: Microtubule-associated protein tau: a component of Alzheimer's paired helical filaments. J Biol Chem 261: 6084–6089, 1986

    Google Scholar 

  4. Grundke-Iqbal I, Iqbal K, Tung Y-C, Quinlan M, Wisniewski HM, Binder LI: Abnormal phosphorylation of the microtubule-associated protein tau in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci USA 83: 4913–4917, 1986

    Google Scholar 

  5. Iqbal K, Grundke-Iqbal I, Smith AJ, George L, Tung Y-C, Zaidi T: Identification and localization of a tau peptide to paired helical filaments of Alzheimer disease. Proc Natl Acad Sci USA 86: 5646–5650, 1989

    Google Scholar 

  6. Brion JP, Hanger DP, Bruce MT, Couck AM, Flament-Durant J, Anderton BT: Tau in Alzheimer's neurofibrillary tangles. Biochem J 273: 127–133, 1991

    Google Scholar 

  7. Morishima-Kawashima M, Hasegawa M, Takio K, Suzuki M, Yoshida H, Titani K, Ihara Y: Proline-directed and non-proline directed phosphorylation of PHF-tau. J Biol Chem 270: 823–829, 1995

    Google Scholar 

  8. Drewes G, Lichtenberg-Kraag B, Coring F, Mandelkow EM, Biernat J, Goris J, Doree M, Mandelkow E: Mitogen-activated protein (MAP) kinase transforms tau protein into an Alzheimer-like state. EMBO J 11: 2131–2138, 1992

    Google Scholar 

  9. Goedert M, Cohen ES, Jakes R, Cohen P: P42 map kinase phosphorylation sites in microtubule-associated protein tau are dephosphorylated by protein phosphatase 2A. FEBS Lett 312: 95–99, 1992

    Google Scholar 

  10. Roder HM, Eden PA, Ingram VM: Brain protein kinase PK40erk converts tau into a PHF-like form as found in Alzheimer's disease. Biochem Biophys Res Commun 193: 639–647, 1993

    Google Scholar 

  11. Vulliet R, Halloran SM, Braun RK, Smith AJ, Lee G: Proline-directed phosphorylation of human tau protein. J Biol Chem 267: 22570–22574, 1992

    Google Scholar 

  12. Baumann K, Mandelkow EM, Biernat J, Piwnica-Worms H, Mandelkow E: Abnormal Alzheimer-like phosphorylation of tau protein by cyclin dependent kinases cdk2 and cdk5. FEBS Lett 336: 417–424, 1993

    Google Scholar 

  13. Paudel HK, Lew J, Ali Z, Wang JH: Brain proline-directed protein kinase phosphorylates tau on sites that are abnormally phosphorylated in tau associated with Alzheimer's paired helical filaments. J Biol Chem 268: 23512–23518, 1993

    Google Scholar 

  14. Mandelkow EM, Drewes G, Biernat J, Gustke N, Van Lint J, Vandenheede JR, Mandelkow E: Glycogen synthase kinase-3 and the Alzheimer-like state of microtubule-associated protein tau. FEBS Lett 314: 315–321, 1992

    Google Scholar 

  15. Ishiguro K, Shiratsuchi A, Sato S, Omori A, Arioka M, Kobayashi S, Uchida T, lmahori K: Glycogen synthase kinase 3β is identical to tau protein kinase I, generating several epitopes of paired helical filaments. FEBS Lett 325: 167–172, 1993

    Google Scholar 

  16. Yang S-D, Song J-S, Yu J-S, Shiah S-G: Protein kinase FA/GSK-3 phosphorylates τ on ser 235-pro and ser 404-pro that are abnormally phosphorylated in Alzheimer's disease brain. J Neurochem 61: 1742–1747, 1993

    Google Scholar 

  17. Singh TJ, Haque N, Grundke-Iqbal I, Iqbal K: Rapid Alzheimer-like phosphorylation of tau by the synergistic actions of non-proline-dependent protein kinases and GSK-3. FEBS Lett 358: 267–272, 1995

    Google Scholar 

  18. Singh TJ, Zaidi T, Grundke-Iqbal I, Iqbal K: Modulation of GSK-3-catalyzed phosphorylation of microtubule-associated protein tau by non-proline-dependent protein kinases. FEBS Lett 358: 4–8, 1995

    Google Scholar 

  19. Litersky JM, Johnson GVW: Phosphorylation of cAMP-dependent protein kinase inhibits the degradation of tau by calpain. J Biol Chem 267: 1563–1568, 1992

    Google Scholar 

  20. Scott CW, Spreen RC, Herman JL, Chow FP, Davison MD, Young J, Caputo CB: Phosphorylation of recombinant tau by cAMP-dependent protein kinase. J Biol Chem 268: 1166–1173, 1993

    Google Scholar 

  21. Singh TJ, Grundke-Iqbal I, Iqbal K: Phosphorylation of tau protein by casein kinase-1 converts it to an abnormal Alzheimer-like state. J Neurochem 64: 1420–1423, 1995

    Google Scholar 

  22. Baudier J, Lee S-H, Cole RD: Separation of the different microtubule-associated tau protein species from bovine brain and their mode II phosphorylation by calcium/phospholipid-dependent protein kinase. J Biol Chem 262: 17584–17590, 1987

    Google Scholar 

  23. Correas I, Diaz-Nido J, Avila J: Microtubule-associated protein tau is phosphorylated by protein kinase C on its tubulin-binding domain. J Biol Chem 267: 15721–15728, 1992

    Google Scholar 

  24. Baudier J, Cole RD: Phosphorylation of tau proteins to a state like that in Alzheimer's brain is catalyzed by calcium/calmodulin-dependent protein kinase and modulated by phospholipids. J Biol Chem 262: 17577–17583, 1991

    Google Scholar 

  25. Singh TJ, Grundke-Iqbal I, McDonald B, Iqbal K: Comparison of the phosphorylation of microtubule-associated protein tau by nonproline-dependent protein kinases. Mol Cell Biochem 131: 181–189, 1994

    Google Scholar 

  26. Greenwood JA, Scott CW, Spreen RC, Caputo CB, Johnson GVW: Casein kinase II preferentially phosphorylates human tau isoforms containing an amino-terminal insert. J Biol Chem 269: 4373–4380, 1994

    Google Scholar 

  27. Greenberg SG, Davies P: A preparation of Alzheimer's paired helical filaments that displays distinct τ proteins by polyacrylamide gel electrophoresis. Proc Natl Acad Sci USA 87: 5827–5831, 1990

    Google Scholar 

  28. Lee VM-Y, Balin BJ, Otvos, Jr L, Trojanowski JQ: A68: a major subunit of paired helical filaments and derivatized forms of normal tau. Science 251: 675–678, 1991

    Google Scholar 

  29. Hanger DP, Hughes K, Woodgett JR, Brion JP, Anderton BH: Glycogen synthase kinase 3 induces Alzheimer's disease-like phosphorylation of tau: generation of paired helical filament epitopes and neuronal localization of the kinase. Neurosci Lett 147: 58–62, 1992

    Google Scholar 

  30. Lichtenberg-Kraag B, Mandelkow EM, Biernat J, Steiner B, Schroter C, Gustke N, Meyer HE, Mandelkow E: Phosphorylation-dependent epitopes of neurofilament antibodies on tau protein and relationship with Alzheimer tau. Proc Natl Acad Sci USA 89: 5384–5388, 1992

    Google Scholar 

  31. Hasegawa M, Watanabe A, Takio K, Suzuki M, Arai T, Titani K, Ihara Y: Characterization of two distinct monoclonal antibodies to paired helical filaments: Further evidence for fetal-type phosphorylation of the tau in paired helical f filaments. J Neurochem 60: 2068–2077, 1993

    Google Scholar 

  32. Biernat J, Mandelkow EM, Schroter C, Lichtenberg-Kraag B, Steiner B, Berling B, Meyer H, Mercken M, Vandermeeren A, Goedert M, Mandelkow E: 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 11: 1593–1597, 1992

    Google Scholar 

  33. Goedert M, Spillantini MG, Jakes R, Rutherford D, Crowther RA: Multiple isoforms of human microtubule-associated protein tau: Sequences and localization in neurofibrillary tangles of Alzheimer's disease. Neuron 3: 519–526, 1989

    Google Scholar 

  34. Singh TJ: Polyamines stimulate the activity of glycogen synthase (casein) kinase-1 from bovine kidney and different rat tissues. Arch Biochem Biophys 267: 167–175, 1988

    Google Scholar 

  35. Tung HYL, Reed LJ: Purification and characterization of protein phosphatase 1 activating kinase from bovine brain cytosol and particulate fractions. J Biol Chem 264: 2985–2990, 1989

    Google Scholar 

  36. Ohmstede C-A, Jensen KJ, Sahyoun NE: Ca2+/calmodulin-dependent protein kinase enriched in cerebellar granule cells. J Biol Chem 264: 5866–5875, 1989

    Google Scholar 

  37. Huang K-P, Chan K-FJ, Singh TJ, Nakabayashi H, Huang FL: Autophosphorylation of rat brain Ca2+-activated and phospholipid-dependent protein kinase. J Biol Chem 261: 12134–12140, 1986

    Google Scholar 

  38. Huang, K-P, Robinson JC: A rapid and sensitive assay method for protein kinase. Anal Biochem 72: 593–599, 1976

    Google Scholar 

  39. Vincent IJ, Davies P: A protein kinase associated with paired helical filaments in Alzheimer disease. Proc Natl Acad Sci USA 89: 2878–2882, 1992

    Google Scholar 

  40. Flotow H, Graves PR, Wang A, Fiol CJ, Roeske RW, Roach PJ: Phosphate groups as substrate determinants for casein kinase I action. J Biol Chem 265: 14264–14269, 1990

    Google Scholar 

  41. Plyte SE, Hughes K, Nikolakaki E, Pulverer BJ, Woodgett JR: Glycogen synthase kinase-3: Functions in oncogenesis and development. Biophys Acta 1114: 147–162, 1992

    Google Scholar 

  42. Liu W-K, Moore WT, Williams RT, Hall FL, Yen S-H: Application of synthetic phospho- and unphospho-peptides to identify phosphorylation sites in a subregion of the tau molecule which is modified in Alzheimer's disease. J Neurosci Res 34: 371–376, 1993

    Google Scholar 

  43. Szendrei GI, Lee VM-Y, Otvos, Jr L: Recognition of the minimal epitope of monoclonal antibody tau-1 depends upon the presence of a phosphate group but not its location. J Neurosci Res 34: 243–249, 1993

    Google Scholar 

  44. Himmler A: Structure of the bovine tau gene: Alternatively spliced transcripts generate a protein family. Mol Cell Biol 9: 1389–1396, 1989

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. New York State Institute for Basic Research in Developmental Disabilities, 10314, Staten Island, New York, USA

    Toolsee J. Singh, Tanweer Zaidi, Inge Grundke-Iqbal & Khalid Iqbal

Authors
  1. Toolsee J. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tanweer Zaidi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Inge Grundke-Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Khalid Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Singh, T.J., Zaidi, T., Grundke-Iqbal, I. et al. Non-proline-dependent protein kinases phosphorylate several sites found in tau from Alzheimer disease brain. Mol Cell Biochem 154, 143–151 (1996). https://doi.org/10.1007/BF00226782

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00226782

Key words

  • GSK-3
  • tau protein
  • protein kinases
  • Alzheimer disease
  • paired helical filaments
  • microtubules