Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration

Nature volume 402, pages 615–622 (1999)Cite this article

Abstract

Cyclin-dependent kinase 5 (Cdk5) is required for proper development of the mammalian central nervous system. To be activated, Cdk5 has to associate with its regulatory subunit, p35. We have found that p25, a truncated form of p35, accumulates in neurons in the brains of patients with Alzheimer's disease. This accumulation correlates with an increase in Cdk5 kinase activity. Unlike p35, p25 is not readily degraded, and binding of p25 to Cdk5 constitutively activates Cdk5, changes its cellular location and alters its substrate specificity. In vivo the p25/Cdk5 complex hyperphosphorylates tau, which reduces tau's ability to associate with microtubules. Moreover, expression of the p25/Cdk5 complex in cultured primary neurons induces cytoskeletal disruption, morphological degeneration and apoptosis. These findings indicate that cleavage of p35, followed by accumulation of p25, may be involved in the pathogenesis of cytoskeletal abnormalities and neuronal death in neurodegenerative diseases.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Accumulation of p25 in brain lysates from patients with AD.
Figure 2: Immunohistochemistry of normal control and AD hippocampal sections.
Figure 3: a, Myristoylation sequence within p35 homologues is conserved.
Figure 4: Western blots of lysates from cycloheximide treatment half-life experiments of p35 and p25 (a) and the p35 G2A mutant (b) in transfected COS-7 cells.
Figure 5: Comparison of tau phosphorylation by p35/Cdk5 and p25/Cdk5.
Figure 6: Effects of the p25/Cdk5 kinase on the neuronal cytoskeleton.
Figure 7: The p25/Cdk5 kinase induces cytoskeletal collapse and nuclear fragmentation.
Figure 8: Quantification of nuclear fragmentation induced by the p25/Cdk5 kinase.

Similar content being viewed by others

References

  1. 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. 267, 25922–25926 (1992).

    CAS  PubMed  Google Scholar 

  2. Meyerson,M. et al. A family of human cdc2-related protein kinases. EMBO J. 11, 2909–2917 (1992).

    Article  CAS  Google Scholar 

  3. Lew,J. et al. A brain-specific activator of cyclin-dependent kinase 5. Nature 371, 423–426 (1994).

    Article  ADS  CAS  Google Scholar 

  4. Ishiguro,K. et al. Identification of the 23 kDa subunit of tau protein kinase II as a putative activator of Cdk5 in bovine brain. FEBS Lett. 342, 203–208 (1994).

    Article  CAS  Google Scholar 

  5. Tsai,L. H., Delalle,I., Caviness,V. S. Jr, Chae,T. & Harlow,E. p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5. Nature 371, 419–423 (1994).

    Article  ADS  CAS  Google Scholar 

  6. Nikolic,M., Dudek,H., Kwon,Y. T., Ramos,Y. F. & Tsai,L. H. The Cdk5/p35 kinase is essential for neurite outgrowth during neuronal differentiation. Genes Dev. 10, 816–825 (1996).

    Article  CAS  Google Scholar 

  7. Paglini,G. et al. Evidence for the participation of the neuron-specific CDK5 activator P35 during laminin-enhanced axonal growth. J. Neurosci. 18, 9858–9869 (1998).

    Article  CAS  Google Scholar 

  8. Kwon,Y. T., Tsai,L. H. & Crandall,J. E. Callosal axon guidance defects in p35–/– mice. J. Comp. Neurol 415, 218–229 (1999).

    Article  CAS  Google Scholar 

  9. Kwon,Y. T. & Tsai,L. H. A novel disruption of cortical development in p35(–/–) mice distinct from reeler. J. Comp. Neurol. 395, 510–522 (1998).

    Article  CAS  Google Scholar 

  10. Chae,T. et al. Mice lacking p35, a neuronal specific activator of Cdk5, display cortical lamination defects, seizures, and adult lethality. Neuron 18, 29–42 (1997).

    Article  CAS  Google Scholar 

  11. Ohshima,T. et al. Targeted disruption of the cyclin-dependent kinase 5 gene results in abnormal corticogenesis, neuronal pathology and perinatal death. Proc. Natl Acad. Sci. USA 93, 11173–11178 (1996).

    Article  ADS  CAS  Google Scholar 

  12. Gilmore,E. C., Ohshima,T., Goffinet,A. M., Kulkarni,A. B. & Herrup,K. Cyclin-dependent kinase 5-deficient mice demonstrate novel developmental arrest in cerebral cortex. J. Neurosci. 18, 6370–6377 (1998).

    Article  CAS  Google Scholar 

  13. Bibb,J. A. et al. Phosphorylation of DARPP-32 by Cdk5 modulates dopamine signalling in neurons. Nature 402, 669–671 (1999).

    Article  ADS  CAS  Google Scholar 

  14. Fletcher,A. I. et al. Regulation of exocytosis by cyclin-dependent kinase 5 via phosphorylation of Munc18. J. Biol. Chem. 274, 4027–4035 (1999).

    Article  CAS  Google Scholar 

  15. Mandelkow,E. M. & Mandelkow,E. Tau in Alzheimer's disease. Trends Cell Biol. 8, 425–427 (1998).

    Article  CAS  Google Scholar 

  16. Spillantini,M. G. & Goedert,M. Tau protein pathology in neurodegenerative diseases. Trends Neurosci. 21, 428–433 (1998).

    Article  CAS  Google Scholar 

  17. Uchida,T. et al. Precursor of Cdk5 activator, the 23 kDa subunit of tau protein kinase II: its sequence and developmental change in brain. FEBS Lett. 355, 35–40 (1994).

    Article  CAS  Google Scholar 

  18. Biernat,J. 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. 11, 1593–1597 (1992).

    Article  CAS  Google Scholar 

  19. Poon,R. Y., Lew,J. & Hunter,T. Identification of functional domains in the neuronal Cdk5 activator protein. J. Biol. Chem. 272, 5703–5708 (1997).

    Article  CAS  Google Scholar 

  20. Nikolic,M., Chou,M. M., Lu,W., Mayer,B. J. & Tsai,L. H. The p35/Cdk5 kinase is a neuron-specific Rac effector that inhibits Pak1 activity. Nature 395, 194–198 (1998).

    Article  ADS  CAS  Google Scholar 

  21. Philpott,A., Porro,E. B., Kirschner,M. W. & Tsai,L. H. The role of cyclin-dependent kinase 5 and a novel regulatory subunit in regulating muscle differentiation and patterning. Genes Dev. 11, 1409–1421 (1997).

    Article  CAS  Google Scholar 

  22. Patrick,G. N., Zhou,P., Kwon,Y. T., Howley,P. M. & Tsai,L. H. p35, the neuronal-specific activator of cyclin-dependent kinase 5 (Cdk5) is degraded by the ubiquitin-proteasome pathway. J. Biol. Chem. 273, 24057–24064 (1998).

    Article  CAS  Google Scholar 

  23. Imahori,K. et al. Possible role of tau protein kinases in pathogenesis of Alzheimer's disease. Neurobiol. Aging 19, S93–98 (1998).

    Article  CAS  Google Scholar 

  24. 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. 336, 417–424 (1993).

    Article  CAS  Google Scholar 

  25. Michel,G. et al. Characterization of tau phosphorylation in glycogen synthase kinase-3β and cyclin dependent kinase-5 activator (p23) transfected cells. Biochim. Biophys. Acta 1380, 177–182 (1998).

    Article  CAS  Google Scholar 

  26. Paudel,H. K. Phosphorylation by neuronal cdc2-like protein kinase promotes dimerization of Tau protein in vitro. J. Biol. Chem. 272, 28328–28334 (1997).

    Article  CAS  Google Scholar 

  27. Paudel,H. K., Lew,J., Ali,Z. & Wang,J. H. 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).

    CAS  PubMed  Google Scholar 

  28. Lovestone,S. et al. Alzheimer's disease-like phosphorylation of the microtubule-associated protein tau by glycogen synthase kinase-3 in transfected mammalian cells. Curr. Biol. 4, 1077–1086 (1994).

    Article  CAS  Google Scholar 

  29. Lovestone,S. & Reynolds,C. H. The phosphorylation of tau: a critical stage in neurodevelopment and neurodegenerative processes. Neuroscience 78, 309–324 (1997).

    Article  CAS  Google Scholar 

  30. Pant Veeranna,A. C., Pant,H. C. & Amin,N. Phosphorylation of human high molecular weight neurofilament protein (hNF-H) by neuronal cyclin-dependent kinase 5 (Cdk5). Brain Res. 765, 259–266 (1997).

    Article  Google Scholar 

  31. Sun,D., Leung,C. L. & Liem,R. K. H. Phosphorylation of the high molecular weight neurofilament protein (NF-H) by Cdk5 and p35. J. Biol. Chem. 271, 14245–14251 (1996).

    Article  CAS  Google Scholar 

  32. Guidato,S., Tsai,L. H., Woodgett,J. & Miller,C. C. Differential cellular phosphorylation of neurofilament heavy side-arms by glycogen synthase kinase-3 and cyclin-dependent kinase-5. J. Neurochem. 66, 1698–1706 (1996).

    Article  CAS  Google Scholar 

  33. Mandelkow,E. M. et al. Tau domains, phosphorylation, and interactions with microtubules. Neurobiol. Aging 16, 355–362; discussion 362–353 (1995).

    Article  CAS  Google Scholar 

  34. Lee,K. Y., Rosales,J. L., Tang,D. & Wang,J. H. Interaction of cyclin-dependent kinase 5 (Cdk5) and neuronal Cdk5 activator in bovine brain. J. Biol. Chem. 271, 1538–1543 (1996).

    Article  CAS  Google Scholar 

  35. Ahuja,H. S., Zhu,Y. & Zakeri,Z. Association of cyclin-dependent kinase 5 and its activator p35 with apoptotic cell death. Dev. Genet. 21, 258–267 (1997).

    Article  CAS  Google Scholar 

  36. Henchcliffe,C. & Burke,R. E. Increased expression of cyclin-dependent kinase 5 in induced apoptotic neuron death in rat substantia nigra. Neurosci. Lett. 230, 41–44 (1997).

    Article  CAS  Google Scholar 

  37. Shirvan,A. et al. Expression of cell cycle-related genes during neuronal apoptosis: is there a distinct pattern. Neurochem. Res. 23, 767–777 (1998).

    Article  CAS  Google Scholar 

  38. Zhang,Q., Ahuja,H. S., Zakeri,Z. F. & Wolgemuth,D. J. Cyclin-dependent kinase 5 is associated with apoptotic cell death during development and tissue remodeling. Dev. Biol. 183, 222–233 (1997).

    Article  CAS  Google Scholar 

  39. Matsubara,M. et al. Site-specific phosphorylation of synapsin I by mitogen-activated protein kinase and Cdk5 and its effects on physiological functions. J. Biol. Chem. 271, 21108–21113 (1996).

    Article  CAS  Google Scholar 

  40. Shuang,R. et al. Regulation of Munc-18/syntaxin 1A interaction by cyclin-dependent kinase 5 in nerve endings. J. Biol. Chem. 273, 4957–4966 (1998).

    Article  CAS  Google Scholar 

  41. Tsai,L. H., Takahashi,T., Caviness,V. S. Jr & Harlow,E. Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system. Development 119, 1029–1040 (1993).

    CAS  PubMed  Google Scholar 

  42. Binder,L. I., Frankfurter,A. & Rebhun,L. I. The distribution of tau in the mammalian central nervous system. J. Cell Biol. 101, 1371–1378 (1985).

    Article  CAS  Google Scholar 

  43. Greenberg,S. G., Davies,P., Schein,J. D. & Binder,L. I. Hydrofluoric acid-treated tau PHF proteins display the same biochemical properties as normal tau. J. Biol. Chem. 267, 564–569 (1992).

    CAS  PubMed  Google Scholar 

  44. Wolozin,B. L., Pruchnicki,A., Dickson,D. W. & Davies,P. A neuronal antigen in the brains of Alzheimer patients. Science 232, 648–650 (1986).

    Article  ADS  CAS  Google Scholar 

  45. Bursztajn,S. et al. Overexpression in neurons of human presenilin-1 or a presenilin-1 familial Alzheimer disease mutant does not enhance apoptosis. J. Neurosci. 18, 9790–9799 (1998).

    Article  CAS  Google Scholar 

  46. Fix,A. S., Ross,J. F., Stitzel,S. R. & Switzer,R. C. Integrated evaluation of central nervous system lesions: stains for neurons, astrocytes, and microglia reveal the spatial and temporal features of MK-801-induced neuronal necrosis in the rat cerebral cortex. Toxicol. Pathol. 24, 291–304 (1996).

    Article  CAS  Google Scholar 

  47. Tanaka,T., Iqbal,K., Trenkner,E., Liu,D. J. & Grundke-Iqbal,I. Abnormally phosphorylated tau in SY5Y human neuroblastoma cells. FEBS Lett. 360, 5–9 (1995).

    Article  CAS  Google Scholar 

  48. Hockenbery,D., Nunez,G., Milliman,C., Schreiber,R. D. & Korsmeyer,S. J. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature 348, 334–336 (1990).

    Article  ADS  CAS  Google Scholar 

  49. Xia,Z., Dickens,M., Raingeaud,J., Davis,R. J. & Greenberg,M. E. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270, 1326–1331 (1995).

    Article  ADS  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Auprin, P. Davies, X. He, R. Neve and J. P. Vonsattel for reagents; Y. Zhou for antibody preparations; D. Smith for help with microscopy; and M. Greenberg, P. Lu, Y. Shi, G. Gill, Y. T. Kwon, D. Smith, V. Tannoch and J. Volker for critical reading of this manuscript. This work was partially supported by NIH grants to L.-H.T. L.-H.T. is an assistant investigator of the Howard Hughes Medical Institute, a Rita Allen Foundation scholar and a recipient of an Ester A. and Joseph Klingenstein Fund award.

Author information

Author notes

  1. Margareta Nikolic

    Present address: Molecular Neurobiology Group, King's College London, London, SE1 9RT, UK

Authors and Affiliations

  1. Department of Pathology, Harvard Medical School,

    Gentry N. Patrick, Lawrence Zukerberg, Margareta Nikolic & Li-Huei Tsai

  2. Howard Hughes Medical Institute, 200 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Li-Huei Tsai

  3. Department of Pathology, Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Lawrence Zukerberg & Suzanne de la Monte

  4. Massachusetts General Hospital—East Cancer Center, Charlestown, 02119, Massachusetts, USA

    Suzanne de la Monte

  5. Department of Neurology, Children's Hospital, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Pieter Dikkes

Authors
  1. Gentry N. Patrick
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lawrence Zukerberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Margareta Nikolic
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Suzanne de la Monte
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pieter Dikkes
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Li-Huei Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Li-Huei Tsai.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Patrick, G., Zukerberg, L., Nikolic, M. et al. Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration. Nature 402, 615–622 (1999). https://doi.org/10.1038/45159

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/45159

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing