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. 1995 Jan;146(1):228–238.

Detection of a Cdc2-related kinase associated with Alzheimer paired helical filaments.

W K Liu 1, R T Williams 1, F L Hall 1, D W Dickson 1, S H Yen 1
PMCID: PMC1870752  PMID: 7856730

Abstract

By immunocytochemical staining and Western blotting, we detected a Cdc2-related kinase in human brains. The kinase is recognized by antibodies against the carboxyl and the amino termini of p34Cdc2 but is not recognized by antibodies against the PSTAIRE motif. It is slightly smaller than p34Cdc2 in molecular mass (approximately 33 kd). This 33-kd Cdc2-related kinase is present in intracellular neurofibrillary tangles in neurons of elderly humans and in Alzheimer's disease, and it is associated with paired helical filaments (PHF) from Alzheimer's disease brains. Unlike the antibodies to the carboxyl and amino termini of p34Cdc2, antibodies to an abundant brain Cdc2-related kinase PSSLARE/Cdk5 did not immunolabel Alzheimer's disease neurofibrillary lesions. PHF preparations were demonstrated to contain kinases capable of phosphorylating histone H1, PHF-Tau, and a synthetic peptide (VAVVRTPPKSPSSAK). By virtue of its physical association with PHF, the 33-kd Cdc2-related kinase may play a role in transforming normal Tau proteins to PHF-Tau characteristic of Alzheimer's disease.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bancher C., Brunner C., Lassmann H., Budka H., Jellinger K., Wiche G., Seitelberger F., Grundke-Iqbal I., Iqbal K., Wisniewski H. M. Accumulation of abnormally phosphorylated tau precedes the formation of neurofibrillary tangles in Alzheimer's disease. Brain Res. 1989 Jan 16;477(1-2):90–99. doi: 10.1016/0006-8993(89)91396-6. [DOI] [PubMed] [Google Scholar]
  2. Baumann K., Mandelkow E. M., Biernat J., Piwnica-Worms H., Mandelkow E. Abnormal Alzheimer-like phosphorylation of tau-protein by cyclin-dependent kinases cdk2 and cdk5. FEBS Lett. 1993 Dec 28;336(3):417–424. doi: 10.1016/0014-5793(93)80849-p. [DOI] [PubMed] [Google Scholar]
  3. Braak H., Braak E., Grundke-Iqbal I., Iqbal K. Occurrence of neuropil threads in the senile human brain and in Alzheimer's disease: a third location of paired helical filaments outside of neurofibrillary tangles and neuritic plaques. Neurosci Lett. 1986 Apr 24;65(3):351–355. doi: 10.1016/0304-3940(86)90288-0. [DOI] [PubMed] [Google Scholar]
  4. Brion J. P., Hanger D. P., Bruce M. T., Couck A. M., Flament-Durand J., Anderton B. H. Tau in Alzheimer neurofibrillary tangles. N- and C-terminal regions are differentially associated with paired helical filaments and the location of a putative abnormal phosphorylation site. Biochem J. 1991 Jan 1;273(Pt 1):127–133. doi: 10.1042/bj2730127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carbonaro-Hall D., Williams R., Wu L., Warburton D., Zeichner-David M., MacDougall M., Tolo V., Hall F. G1 expression and multistage dynamics of cyclin A in human osteosarcoma cells. Oncogene. 1993 Jun;8(6):1649–1659. [PubMed] [Google Scholar]
  6. Dickson D. W., Ksiezak-Reding H., Liu W. K., Davies P., Crowe A., Yen S. H. Immunocytochemistry of neurofibrillary tangles with antibodies to subregions of tau protein: identification of hidden and cleaved tau epitopes and a new phosphorylation site. Acta Neuropathol. 1992;84(6):596–605. doi: 10.1007/BF00227736. [DOI] [PubMed] [Google Scholar]
  7. Draetta G., Beach D. Activation of cdc2 protein kinase during mitosis in human cells: cell cycle-dependent phosphorylation and subunit rearrangement. Cell. 1988 Jul 1;54(1):17–26. doi: 10.1016/0092-8674(88)90175-4. [DOI] [PubMed] [Google Scholar]
  8. Drewes G., Lichtenberg-Kraag B., Döring F., Mandelkow E. M., Biernat J., Goris J., Dorée M., Mandelkow E. Mitogen activated protein (MAP) kinase transforms tau protein into an Alzheimer-like state. EMBO J. 1992 Jun;11(6):2131–2138. doi: 10.1002/j.1460-2075.1992.tb05272.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elledge S. J., Richman R., Hall F. L., Williams R. T., Lodgson N., Harper J. W. CDK2 encodes a 33-kDa cyclin A-associated protein kinase and is expressed before CDC2 in the cell cycle. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2907–2911. doi: 10.1073/pnas.89.7.2907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Erickson A. K., Payne D. M., Martino P. A., Rossomando A. J., Shabanowitz J., Weber M. J., Hunt D. F., Sturgill T. W. Identification by mass spectrometry of threonine 97 in bovine myelin basic protein as a specific phosphorylation site for mitogen-activated protein kinase. J Biol Chem. 1990 Nov 15;265(32):19728–19735. [PubMed] [Google Scholar]
  11. Fiol C. J., Haseman J. H., Wang Y. H., Roach P. J., Roeske R. W., Kowalczuk M., DePaoli-Roach A. A. Phosphoserine as a recognition determinant for glycogen synthase kinase-3: phosphorylation of a synthetic peptide based on the G-component of protein phosphatase-1. Arch Biochem Biophys. 1988 Dec;267(2):797–802. doi: 10.1016/0003-9861(88)90089-6. [DOI] [PubMed] [Google Scholar]
  12. Fiol C. J., Mahrenholz A. M., Wang Y., Roeske R. W., Roach P. J. Formation of protein kinase recognition sites by covalent modification of the substrate. Molecular mechanism for the synergistic action of casein kinase II and glycogen synthase kinase 3. J Biol Chem. 1987 Oct 15;262(29):14042–14048. [PubMed] [Google Scholar]
  13. Goedert M., Jakes R., Crowther R. A., Six J., Lübke U., Vandermeeren M., Cras P., Trojanowski J. Q., Lee V. M. The abnormal phosphorylation of tau protein at Ser-202 in Alzheimer disease recapitulates phosphorylation during development. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5066–5070. doi: 10.1073/pnas.90.11.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greenberg S. G., Davies P. A preparation of Alzheimer paired helical filaments that displays distinct tau proteins by polyacrylamide gel electrophoresis. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5827–5831. doi: 10.1073/pnas.87.15.5827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grundke-Iqbal I., Iqbal K., Tung Y. C., Quinlan M., Wisniewski H. M., Binder L. I. Abnormal phosphorylation of the microtubule-associated protein tau (tau) in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4913–4917. doi: 10.1073/pnas.83.13.4913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hall F. L., Braun R. K., Mitchell J. P., Vulliet P. R. Phosphorylation of cytoskeletal proteins by proline directed protein kinase. Proc West Pharmacol Soc. 1990;33:213–217. [PubMed] [Google Scholar]
  17. Hall F. L., Vulliet P. R. Proline-directed protein phosphorylation and cell cycle regulation. Curr Opin Cell Biol. 1991 Apr;3(2):176–184. doi: 10.1016/0955-0674(91)90136-m. [DOI] [PubMed] [Google Scholar]
  18. Hall F. L., Williams R. T., Wu L., Wu F., Carbonaro-Hall D. A., Harper J. W., Warburton D. Two potentially oncogenic cyclins, cyclin A and cyclin D1, share common properties of subunit configuration, tyrosine phosphorylation and physical association with the Rb protein. Oncogene. 1993 May;8(5):1377–1384. [PubMed] [Google Scholar]
  19. Hanger D. P., Hughes K., Woodgett J. R., Brion J. P., Anderton B. H. Glycogen synthase kinase-3 induces Alzheimer's disease-like phosphorylation of tau: generation of paired helical filament epitopes and neuronal localisation of the kinase. Neurosci Lett. 1992 Nov 23;147(1):58–62. doi: 10.1016/0304-3940(92)90774-2. [DOI] [PubMed] [Google Scholar]
  20. Hasegawa M., Morishima-Kawashima M., Takio K., Suzuki M., Titani K., Ihara Y. Protein sequence and mass spectrometric analyses of tau in the Alzheimer's disease brain. J Biol Chem. 1992 Aug 25;267(24):17047–17054. [PubMed] [Google Scholar]
  21. Hisanaga S., Ishiguro K., Uchida T., Okumura E., Okano T., Kishimoto T. Tau protein kinase II has a similar characteristic to cdc2 kinase for phosphorylating neurofilament proteins. J Biol Chem. 1993 Jul 15;268(20):15056–15060. [PubMed] [Google Scholar]
  22. Iqbal K., Grundke-Iqbal I., Smith A. J., 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 U S A. 1989 Jul;86(14):5646–5650. doi: 10.1073/pnas.86.14.5646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ishiguro K., Omori A., Sato K., Tomizawa K., Imahori K., Uchida T. A serine/threonine proline kinase activity is included in the tau protein kinase fraction forming a paired helical filament epitope. Neurosci Lett. 1991 Jul 22;128(2):195–198. doi: 10.1016/0304-3940(91)90259-v. [DOI] [PubMed] [Google Scholar]
  24. Kanemaru K., Takio K., Miura R., Titani K., Ihara Y. Fetal-type phosphorylation of the tau in paired helical filaments. J Neurochem. 1992 May;58(5):1667–1675. doi: 10.1111/j.1471-4159.1992.tb10039.x. [DOI] [PubMed] [Google Scholar]
  25. Kenessey A., Yen S. H. The extent of phosphorylation of fetal tau is comparable to that of PHF-tau from Alzheimer paired helical filaments. Brain Res. 1993 Nov 26;629(1):40–46. doi: 10.1016/0006-8993(93)90478-6. [DOI] [PubMed] [Google Scholar]
  26. Kobayashi S., Ishiguro K., Omori A., Takamatsu M., Arioka M., Imahori K., Uchida T. A cdc2-related kinase PSSALRE/cdk5 is homologous with the 30 kDa subunit of tau protein kinase II, a proline-directed protein kinase associated with microtubule. FEBS Lett. 1993 Dec 6;335(2):171–175. doi: 10.1016/0014-5793(93)80723-8. [DOI] [PubMed] [Google Scholar]
  27. Ksiezak-Reding H., Binder L. I., Yen S. H. Alzheimer disease proteins (A68) share epitopes with tau but show distinct biochemical properties. J Neurosci Res. 1990 Mar;25(3):420–430. doi: 10.1002/jnr.490250320. [DOI] [PubMed] [Google Scholar]
  28. Ksiezak-Reding H., Liu W. K., Yen S. H. Phosphate analysis and dephosphorylation of modified tau associated with paired helical filaments. Brain Res. 1992 Dec 4;597(2):209–219. doi: 10.1016/0006-8993(92)91476-u. [DOI] [PubMed] [Google Scholar]
  29. Ksiezak-Reding H., Yen S. H. Structural stability of paired helical filaments requires microtubule-binding domains of tau: a model for self-association. Neuron. 1991 May;6(5):717–728. doi: 10.1016/0896-6273(91)90169-z. [DOI] [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. Lang E., Szendrei G. I., Lee V. M., Otvos L., Jr Immunological and conformation characterization of a phosphorylated immunodominant epitope on the paired helical filaments found in Alzheimer's disease. Biochem Biophys Res Commun. 1992 Sep 16;187(2):783–790. doi: 10.1016/0006-291x(92)91264-q. [DOI] [PubMed] [Google Scholar]
  32. Lee V. M., Balin B. J., Otvos L., Jr, Trojanowski J. Q. A68: a major subunit of paired helical filaments and derivatized forms of normal Tau. Science. 1991 Feb 8;251(4994):675–678. doi: 10.1126/science.1899488. [DOI] [PubMed] [Google Scholar]
  33. Lew J., Winkfein R. J., Paudel H. K., Wang J. H. Brain proline-directed protein kinase is a neurofilament kinase which displays high sequence homology to p34cdc2. J Biol Chem. 1992 Dec 25;267(36):25922–25926. [PubMed] [Google Scholar]
  34. Liu W. K., Dickson D. W., Yen S. H. Heterogeneity of tau proteins in Alzheimer's disease. Evidence for increased expression of an isoform and preferential distribution of a phosphorylated isoform in neurites. Am J Pathol. 1993 Feb;142(2):387–394. [PMC free article] [PubMed] [Google Scholar]
  35. Liu W. K., Ksiezak-Reding H., Yen S. H. Abnormal tau proteins from Alzheimer's disease brains. Purification and amino acid analysis. J Biol Chem. 1991 Nov 15;266(32):21723–21727. [PubMed] [Google Scholar]
  36. Liu W. K., Moore W. T., Williams R. T., Hall F. L., 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. 1993 Feb 15;34(3):371–376. doi: 10.1002/jnr.490340315. [DOI] [PubMed] [Google Scholar]
  37. Mandelkow E. M., Drewes G., Biernat J., Gustke N., Van Lint J., Vandenheede J. R., Mandelkow E. Glycogen synthase kinase-3 and the Alzheimer-like state of microtubule-associated protein tau. FEBS Lett. 1992 Dec 21;314(3):315–321. doi: 10.1016/0014-5793(92)81496-9. [DOI] [PubMed] [Google Scholar]
  38. Mawal-Dewan M., Sen P. C., Abdel-Ghany M., Shalloway D., Racker E. Phosphorylation of tau protein by purified p34cdc28 and a related protein kinase from neurofilaments. J Biol Chem. 1992 Sep 25;267(27):19705–19709. [PubMed] [Google Scholar]
  39. Meyerson M., Enders G. H., Wu C. L., Su L. K., Gorka C., Nelson C., Harlow E., Tsai L. H. A family of human cdc2-related protein kinases. EMBO J. 1992 Aug;11(8):2909–2917. doi: 10.1002/j.1460-2075.1992.tb05360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ray L. B., Sturgill T. W. Characterization of insulin-stimulated microtubule-associated protein kinase. Rapid isolation and stabilization of a novel serine/threonine kinase from 3T3-L1 cells. J Biol Chem. 1988 Sep 5;263(25):12721–12727. [PubMed] [Google Scholar]
  41. Roder H. M., Eden P. A., Ingram V. M. Brain protein kinase PK40erk converts TAU into a PHF-like form as found in Alzheimer's disease. Biochem Biophys Res Commun. 1993 Jun 15;193(2):639–647. doi: 10.1006/bbrc.1993.1672. [DOI] [PubMed] [Google Scholar]
  42. Rossomando A. J., Sanghera J. S., Marsden L. A., Weber M. J., Pelech S. L., Sturgill T. W. Biochemical characterization of a family of serine/threonine protein kinases regulated by tyrosine and serine/threonine phosphorylations. J Biol Chem. 1991 Oct 25;266(30):20270–20275. [PubMed] [Google Scholar]
  43. Scott C. W., Vulliet P. R., Caputo C. B. Phosphorylation of tau by proline-directed protein kinase (p34cdc2/p58cyclin A) decreases tau-induced microtubule assembly and antibody SMI33 reactivity. Brain Res. 1993 May 21;611(2):237–242. doi: 10.1016/0006-8993(93)90508-k. [DOI] [PubMed] [Google Scholar]
  44. Shetty K. T., Link W. T., Pant H. C. cdc2-like kinase from rat spinal cord specifically phosphorylates KSPXK motifs in neurofilament proteins: isolation and characterization. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6844–6848. doi: 10.1073/pnas.90.14.6844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Trojanowski J. Q., Mawal-Dewan M., Schmidt M. L., Martin J., Lee V. M. Localization of the mitogen activated protein kinase ERK2 in Alzheimer's disease neurofibrillary tangles and senile plaque neurites. Brain Res. 1993 Aug 6;618(2):333–337. doi: 10.1016/0006-8993(93)91286-2. [DOI] [PubMed] [Google Scholar]
  46. Vincent I. J., Davies P. A protein kinase associated with paired helical filaments in Alzheimer disease. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2878–2882. doi: 10.1073/pnas.89.7.2878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vulliet P. R., Hall F. L., Mitchell J. P., Hardie D. G. Identification of a novel proline-directed serine/threonine protein kinase in rat pheochromocytoma. J Biol Chem. 1989 Sep 25;264(27):16292–16298. [PubMed] [Google Scholar]
  48. Vulliet R., Halloran S. M., Braun R. K., Smith A. J., Lee G. Proline-directed phosphorylation of human Tau protein. J Biol Chem. 1992 Nov 5;267(31):22570–22574. [PubMed] [Google Scholar]
  49. Williams R. T., Carbonaro-Hall D. A., Hall F. L. Co-purification of p34cdc2/p58cyclin A proline-directed protein kinase and the retinoblastoma tumor susceptibility gene product: interaction of an oncogenic serine/threonine protein kinase with a tumor-suppressor protein. Oncogene. 1992 Mar;7(3):423–432. [PubMed] [Google Scholar]

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