Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1996 February 1; 132(3): 413–425.
PMCID: PMC2120731

Mitotic mechanisms in Alzheimer's disease?


The mechanism(s) leading to widespread hyper-phosphorylation of proteins in Alzheimer's disease (AD) are unknown. We have characterized seven new monoclonal antibodies recognizing independent phospho- epitopes in the paired helical filament proteins (PHF) found in AD brain. These antibodies show pronounced immunoreactivity with cultured human neuroblastoma cells that are in the M phase of cell division, but have no discernible reactivity with interphase cells. Immunoreactivity with these antibodies does not localize to the microtubule spindles or chromosomes in M phase, but is confined to the surrounding cytoplasm. Similar staining in M phase is observed with cultured cells of various tissue types and species. Cells arrested in M phase with the microtubule depolymerizing agent, nocodazole, show marked increases in immunoreactivity with the antibodies by immunofluorescence staining, ELISA, and immunoblotting. In neuroblastoma cells, the appearance of the TG/MC phospho-epitopes coincides with activation of mitotic protein kinases, but not with the activity of the neuronal specific cyclin- dependent kinase, cdk5. These data suggest that the TG/MC epitopes are conserved mitotic phospho-epitopes produced as a result of increased mitotic kinase activity. To investigate this possibility in AD, we examined the staining of human brain tissue with MPM-2, a marker antibody for mitotic phospho-epitopes. It was found that MPM-2 reacts strongly with neurofibrillary tangles, neuritic processes, and neurons in AD but has no staining in normal human brain. Our data suggest that accumulation of phospho-epitopes in AD may result from activation of mitotic posttranslational mechanisms which do not normally operate in mature neurons of brain.

Full Text

The Full Text of this article is available as a PDF (9.6M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Azzi L, Meijer L, Ostvold AC, Lew J, Wang JH. Purification of a 15-kDa cdk4- and cdk5-binding protein. J Biol Chem. 1994 May 6;269(18):13279–13288. [PubMed]
  • Baudier J, Cole RD. Phosphorylation of tau proteins to a state like that in Alzheimer's brain is catalyzed by a calcium/calmodulin-dependent kinase and modulated by phospholipids. J Biol Chem. 1987 Dec 25;262(36):17577–17583. [PubMed]
  • 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. 1993 Dec 28;336(3):417–424. [PubMed]
  • Brizuela L, Draetta G, Beach D. p13suc1 acts in the fission yeast cell division cycle as a component of the p34cdc2 protein kinase. EMBO J. 1987 Nov;6(11):3507–3514. [PubMed]
  • Davis FM, Tsao TY, Fowler SK, Rao PN. Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2926–2930. [PubMed]
  • Dickson DW. Apoptosis in the brain. Physiology and pathology. Am J Pathol. 1995 May;146(5):1040–1044. [PubMed]
  • 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. [PubMed]
  • Draetta G, Brizuela L, Potashkin J, Beach D. Identification of p34 and p13, human homologs of the cell cycle regulators of fission yeast encoded by cdc2+ and suc1+. Cell. 1987 Jul 17;50(2):319–325. [PubMed]
  • Drechsel DN, Hyman AA, Cobb MH, Kirschner MW. Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau. Mol Biol Cell. 1992 Oct;3(10):1141–1154. [PMC free article] [PubMed]
  • Drewes G, Lichtenberg-Kraag B, Döring F, Mandelkow EM, 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. [PubMed]
  • Engle DB, Doonan JH, Morris NR. Cell-cycle modulation of MPM-2-specific spindle pole body phosphorylation in Aspergillus nidulans. Cell Motil Cytoskeleton. 1988;10(3):434–437. [PubMed]
  • Freeman RS, Estus S, Johnson EM., Jr Analysis of cell cycle-related gene expression in postmitotic neurons: selective induction of Cyclin D1 during programmed cell death. Neuron. 1994 Feb;12(2):343–355. [PubMed]
  • Goedert M, Spillantini MG, Cairns NJ, Crowther RA. Tau proteins of Alzheimer paired helical filaments: abnormal phosphorylation of all six brain isoforms. Neuron. 1992 Jan;8(1):159–168. [PubMed]
  • Goedert M, Cohen ES, Jakes R, Cohen P. p42 MAP kinase phosphorylation sites in microtubule-associated protein tau are dephosphorylated by protein phosphatase 2A1. Implications for Alzheimer's disease [corrected]. FEBS Lett. 1992 Nov 2;312(1):95–99. [PubMed]
  • Greenberg SG, Davies P, Schein JD, Binder LI. Hydrofluoric acid-treated tau PHF proteins display the same biochemical properties as normal tau. J Biol Chem. 1992 Jan 5;267(1):564–569. [PubMed]
  • Grundke-Iqbal I, Iqbal K, Tung YC, Quinlan M, Wisniewski HM, Binder LI. 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. [PubMed]
  • Haass C, Koo EH, Mellon A, Hung AY, Selkoe DJ. Targeting of cell-surface beta-amyloid precursor protein to lysosomes: alternative processing into amyloid-bearing fragments. Nature. 1992 Jun 11;357(6378):500–503. [PubMed]
  • Hall FL, Vulliet PR. Proline-directed protein phosphorylation and cell cycle regulation. Curr Opin Cell Biol. 1991 Apr;3(2):176–184. [PubMed]
  • Hall FL, Braun RK, Mitchell JP, Vulliet PR. Phosphorylation of cytoskeletal proteins by proline directed protein kinase. Proc West Pharmacol Soc. 1990;33:213–217. [PubMed]
  • Hayes TE, Valtz NL, McKay RD. Downregulation of CDC2 upon terminal differentiation of neurons. New Biol. 1991 Mar;3(3):259–269. [PubMed]
  • Hellmich MR, Pant HC, Wada E, Battey JF. Neuronal cdc2-like kinase: a cdc2-related protein kinase with predominantly neuronal expression. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10867–10871. [PubMed]
  • 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]
  • Hoshi M, Nishida E, Miyata Y, Sakai H, Miyoshi T, Ogawara H, Akiyama T. Protein kinase C phosphorylates tau and induces its functional alterations. FEBS Lett. 1987 Jun 15;217(2):237–241. [PubMed]
  • Ishiguro K, Takamatsu M, Tomizawa K, Omori A, Takahashi M, Arioka M, Uchida T, Imahori K. Tau protein kinase I converts normal tau protein into A68-like component of paired helical filaments. J Biol Chem. 1992 May 25;267(15):10897–10901. [PubMed]
  • Ishiguro K, Kobayashi S, Omori A, Takamatsu M, Yonekura S, Anzai K, Imahori K, Uchida T. Identification of the 23 kDa subunit of tau protein kinase II as a putative activator of cdk5 in bovine brain. FEBS Lett. 1994 Apr 4;342(2):203–208. [PubMed]
  • 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. [PubMed]
  • Kang J, Lemaire HG, Unterbeck A, Salbaum JM, Masters CL, Grzeschik KH, Multhaup G, Beyreuther K, Müller-Hill B. The precursor of Alzheimer's disease amyloid A4 protein resembles a cell-surface receptor. Nature. 1987 Feb 19;325(6106):733–736. [PubMed]
  • Ksiezak-Reding H, Binder LI, Yen SH. Immunochemical and biochemical characterization of tau proteins in normal and Alzheimer's disease brains with Alz 50 and Tau-1. J Biol Chem. 1988 Jun 15;263(17):7948–7953. [PubMed]
  • Ksiezak-Reding H, Liu WK, Yen SH. Phosphate analysis and dephosphorylation of modified tau associated with paired helical filaments. Brain Res. 1992 Dec 4;597(2):209–219. [PubMed]
  • Kuang J, Ashorn CL, Gonzalez-Kuyvenhoven M, Penkala JE. cdc25 is one of the MPM-2 antigens involved in the activation of maturation-promoting factor. Mol Biol Cell. 1994 Feb;5(2):135–145. [PMC free article] [PubMed]
  • Lassmann H, Bancher C, Breitschopf H, Wegiel J, Bobinski M, Jellinger K, Wisniewski HM. Cell death in Alzheimer's disease evaluated by DNA fragmentation in situ. Acta Neuropathol. 1995;89(1):35–41. [PubMed]
  • Ledesma MD, Correas I, Avila J, Díaz-Nido J. Implication of brain cdc2 and MAP2 kinases in the phosphorylation of tau protein in Alzheimer's disease. FEBS Lett. 1992 Aug 17;308(2):218–224. [PubMed]
  • Lee VM, Balin BJ, Otvos L, Jr, Trojanowski JQ. A68: a major subunit of paired helical filaments and derivatized forms of normal Tau. Science. 1991 Feb 8;251(4994):675–678. [PubMed]
  • Lee S, Christakos S, Small MB. Apoptosis and signal transduction: clues to a molecular mechanism. Curr Opin Cell Biol. 1993 Apr;5(2):286–291. [PubMed]
  • Lew J, Winkfein RJ, Paudel HK, Wang JH. 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]
  • Lew J, Huang QQ, Qi Z, Winkfein RJ, Aebersold R, Hunt T, Wang JH. A brain-specific activator of cyclin-dependent kinase 5. Nature. 1994 Sep 29;371(6496):423–426. [PubMed]
  • Lindwall G, Cole RD. Phosphorylation affects the ability of tau protein to promote microtubule assembly. J Biol Chem. 1984 Apr 25;259(8):5301–5305. [PubMed]
  • Litersky JM, Johnson GV. Phosphorylation by cAMP-dependent protein kinase inhibits the degradation of tau by calpain. J Biol Chem. 1992 Jan 25;267(3):1563–1568. [PubMed]
  • Liu WK, Williams RT, Hall FL, Dickson DW, Yen SH. Detection of a Cdc2-related kinase associated with Alzheimer paired helical filaments. Am J Pathol. 1995 Jan;146(1):228–238. [PubMed]
  • Lu Q, Soria JP, Wood JG. p44mpk MAP kinase induces Alzheimer type alterations in tau function and in primary hippocampal neurons. J Neurosci Res. 1993 Jul 1;35(4):439–444. [PubMed]
  • Maller JL. Mitotic control. Curr Opin Cell Biol. 1991 Apr;3(2):269–275. [PubMed]
  • 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. 1992 Dec 21;314(3):315–321. [PubMed]
  • Masliah E, Mallory M, Alford M, Hansen LA, Saitoh T. Immunoreactivity of the nuclear antigen p105 is associated with plaques and tangles in Alzheimer's disease. Lab Invest. 1993 Nov;69(5):562–569. [PubMed]
  • Mawal-Dewan M, Sen PC, 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]
  • Meikrantz W, Gisselbrecht S, Tam SW, Schlegel R. Activation of cyclin A-dependent protein kinases during apoptosis. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3754–3758. [PubMed]
  • Meyerson M, Enders GH, Wu CL, Su LK, Gorka C, Nelson C, Harlow E, Tsai LH. A family of human cdc2-related protein kinases. EMBO J. 1992 Aug;11(8):2909–2917. [PubMed]
  • Nigg EA. Targets of cyclin-dependent protein kinases. Curr Opin Cell Biol. 1993 Apr;5(2):187–193. [PubMed]
  • Norbury C, MacFarlane M, Fearnhead H, Cohen GM. Cdc2 activation is not required for thymocyte apoptosis. Biochem Biophys Res Commun. 1994 Aug 15;202(3):1400–1406. [PubMed]
  • Nurse P. Universal control mechanism regulating onset of M-phase. Nature. 1990 Apr 5;344(6266):503–508. [PubMed]
  • Oberhammer FA, Hochegger K, Fröschl G, Tiefenbacher R, Pavelka M. Chromatin condensation during apoptosis is accompanied by degradation of lamin A+B, without enhanced activation of cdc2 kinase. J Cell Biol. 1994 Aug;126(4):827–837. [PMC free article] [PubMed]
  • 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. 1993 Nov 5;268(31):23512–23518. [PubMed]
  • Pope WB, Lambert MP, Leypold B, Seupaul R, Sletten L, Krafft G, Klein WL. Microtubule-associated protein tau is hyperphosphorylated during mitosis in the human neuroblastoma cell line SH-SY5Y. Exp Neurol. 1994 Apr;126(2):185–194. [PubMed]
  • Shi L, Nishioka WK, Th'ng J, Bradbury EM, Litchfield DW, Greenberg AH. Premature p34cdc2 activation required for apoptosis. Science. 1994 Feb 25;263(5150):1143–1145. [PubMed]
  • Smith TW, Lippa CF. Ki-67 immunoreactivity in Alzheimer's disease and other neurodegenerative disorders. J Neuropathol Exp Neurol. 1995 May;54(3):297–303. [PubMed]
  • Steiner B, Mandelkow EM, Biernat J, Gustke N, Meyer HE, Schmidt B, Mieskes G, Söling HD, Drechsel D, Kirschner MW, et al. Phosphorylation of microtubule-associated protein tau: identification of the site for Ca2(+)-calmodulin dependent kinase and relationship with tau phosphorylation in Alzheimer tangles. EMBO J. 1990 Nov;9(11):3539–3544. [PubMed]
  • Steinmann KE, Belinsky GS, Lee D, Schlegel R. Chemically induced premature mitosis: differential response in rodent and human cells and the relationship to cyclin B synthesis and p34cdc2/cyclin B complex formation. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6843–6847. [PubMed]
  • Suzuki T, Oishi M, Marshak DR, Czernik AJ, Nairn AC, Greengard P. Cell cycle-dependent regulation of the phosphorylation and metabolism of the Alzheimer amyloid precursor protein. EMBO J. 1994 Mar 1;13(5):1114–1122. [PubMed]
  • Tombes RM, Peloquin JG, Borisy GG. Specific association of an M-phase kinase with isolated mitotic spindles and identification of two of its substrates as MAP4 and MAP1B. Cell Regul. 1991 Nov;2(11):861–874. [PMC free article] [PubMed]
  • Tsai LH, Takahashi T, Caviness VS, Jr, Harlow E. Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system. Development. 1993 Dec;119(4):1029–1040. [PubMed]
  • Tsai LH, Delalle I, Caviness VS, Jr, Chae T, Harlow E. p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5. Nature. 1994 Sep 29;371(6496):419–423. [PubMed]
  • Ucker DS. Death by suicide: one way to go in mammalian cellular development? New Biol. 1991 Feb;3(2):103–109. [PubMed]
  • Vandre DD, Davis FM, Rao PN, Borisy GG. Phosphoproteins are components of mitotic microtubule organizing centers. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4439–4443. [PubMed]
  • Vandré DD, Davis FM, Rao PN, Borisy GG. Distribution of cytoskeletal proteins sharing a conserved phosphorylated epitope. Eur J Cell Biol. 1986 Jun;41(1):72–81. [PubMed]
  • Vandré DD, Centonze VE, Peloquin J, Tombes RM, Borisy GG. Proteins of the mammalian mitotic spindle: phosphorylation/dephosphorylation of MAP-4 during mitosis. J Cell Sci. 1991 Apr;98(Pt 4):577–588. [PubMed]
  • Verde F, Labbé JC, Dorée M, Karsenti E. Regulation of microtubule dynamics by cdc2 protein kinase in cell-free extracts of Xenopus eggs. Nature. 1990 Jan 18;343(6255):233–238. [PubMed]
  • Vincent IJ, 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. [PubMed]
  • Vincent I, Rosado M, Kim E, Davies P. Increased production of paired helical filament epitopes in a cell culture system reduces the turnover of tau. J Neurochem. 1994 Feb;62(2):715–723. [PubMed]
  • Williams GT, Smith CA. Molecular regulation of apoptosis: genetic controls on cell death. Cell. 1993 Sep 10;74(5):777–779. [PubMed]
  • Williams GT, Smith CA, McCarthy NJ, Grimes EA. Apoptosis: final control point in cell biology. Trends Cell Biol. 1992 Sep;2(9):263–267. [PubMed]
  • Wolozin BL, Pruchnicki A, Dickson DW, Davies P. A neuronal antigen in the brains of Alzheimer patients. Science. 1986 May 2;232(4750):648–650. [PubMed]
  • Wood JG, Mirra SS, Pollock NJ, Binder LI. Neurofibrillary tangles of Alzheimer disease share antigenic determinants with the axonal microtubule-associated protein tau (tau) Proc Natl Acad Sci U S A. 1986 Jun;83(11):4040–4043. [PubMed]
  • Wood JG, Lu Q, Reich C, Zinsmeister P. Proline-directed kinase systems in Alzheimer's disease pathology. Neurosci Lett. 1993 Jun 25;156(1-2):83–86. [PubMed]
  • Wordeman L, Davis FM, Rao PN, Cande WZ. Distribution of phosphorylated spindle-associated proteins in the diatom Stephanopyxis turris. Cell Motil Cytoskeleton. 1989;12(1):33–41. [PubMed]
  • Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach D. p21 is a universal inhibitor of cyclin kinases. Nature. 1993 Dec 16;366(6456):701–704. [PubMed]
  • Yamashita K, Yasuda H, Pines J, Yasumoto K, Nishitani H, Ohtsubo M, Hunter T, Sugimura T, Nishimoto T. Okadaic acid, a potent inhibitor of type 1 and type 2A protein phosphatases, activates cdc2/H1 kinase and transiently induces a premature mitosis-like state in BHK21 cells. EMBO J. 1990 Dec;9(13):4331–4338. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press