Skip to main content
Download PDF
  • Short Communication
  • Published:

p600 stabilizes microtubules to prevent the aggregation of CaMKIIα during photoconductive stimulation

Cellular & Molecular Biology Letters volume 19, pages 381–392 (2014)Cite this article

Abstract

The large microtubule-associated/Ca2+-signalling protein p600 (also known as UBR4) is required for hippocampal neuronal survival upon Ca2+ dyshomeostasis induced by glutamate treatment. During this process, p600 prevents aggregation of the Ca2+/calmodulin-dependent kinase IIα (CaMKIIα), a proxy of neuronal death, via direct binding to calmodulin in a microtubuleindependent manner. Using photoconductive stimulation coupled with live imaging of single neurons, we identified a distinct mechanism of prevention of CaMKIIα aggregation by p600. Upon direct depolarization, CaMKIIα translocates to microtubules. In the absence of p600, this translocation is interrupted in favour of a sustained self-aggregation that is prevented by the microtubule-stabilizing drug paclitaxel. Thus, during photoconductive stimulation, p600 prevents the aggregation of CaMKIIα by stabilizing microtubules. The effectiveness of this stabilization for preventing CaMKIIα aggregation during direct depolarization but not during glutamate treatment suggests a model wherein p600 has two modes of action depending on the source of cytosolic Ca2+.

Abbreviations

CaM:

calmodulin

CaMKII:

Ca2+/calmodulin-dependent kinase II

GFP:

green fluorescent protein

NMDA:

N-methyl-D-aspartate

AMPA:

α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid

DMSO:

dimethyl sulfoxide

References

  1. Erondu, E. and Kennedy, B. Regional distribution of type II Ca2+/Calmodulindependent protein kinase in rat brain. J. Neurosci. 5 (1985) 3270–3277.

    PubMed  CAS  Google Scholar 

  2. Lisman, J., Schulman, H. and Cline, H. The molecular basis of CaMKII function in synaptic and behavioural memory. Nat. Rev. Neurosci. 3 (2002) 175–190.

    Article  PubMed  CAS  Google Scholar 

  3. Kochhar, A., Saitoh, T. and Zivin, J. Spinal cord ischemia reduces calcium/calmodulin-dependent protein kinase activity. Brain Res. 542 (1991) 141–146.

    Article  PubMed  CAS  Google Scholar 

  4. Aronowski, J., Grotta, J. and Waxham, M. Ischemia-induced translocation of Ca2+/calmodulin-dependent protein kinase II: potential role in neuronal damage. J. Neurochem. 58 (1992) 1743–1753.

    Article  PubMed  CAS  Google Scholar 

  5. Hanson, S., Grotta, J., Waxham, M., Aronowski, J. and Ostrow, P. Calcium/calmodulin-dependent protein kinase II activity in focal ischemia with reperfusion in rats. Stroke 25 (1994) 466–473.

    Article  PubMed  CAS  Google Scholar 

  6. Hudmon, A., Aronowski, J., Kolb, S. and Waxham, M. Inactivation and selfassociation of Ca2+/calmodulin-dependent protein kinase II during autophosphorylation. J. Biol. Chem. 271 (1996) 8800–8808.

    Article  PubMed  CAS  Google Scholar 

  7. Dosemeci, A., Reese, T., Petersen, J. and Tao-Cheng, J. A novel particulate form of Ca2+/CaMKII-dependent protein kinase II in neurons. J. Neurosci. 20 (2000) 3076–3084.

    PubMed  CAS  Google Scholar 

  8. Hudmon, A., Lebel, E., Roy, H., Sik, A., Schulman, H., Waxham, M. and De Koninck, P. A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association. J. Neurosci. 25 (2005) 6971–6983.

    Article  PubMed  CAS  Google Scholar 

  9. Tao-Cheng, J., Vinade, L., Smith, C., Winters, C., Ward, R., Brightman, M., Reese, T. and Dosemeci, A. Sustained elevation of calcium induces Ca2+/calmodulin-dependent protein kinase II clusters in hippocampal neurons. Neuroscience 106 (2001) 69–78.

    Article  PubMed  CAS  Google Scholar 

  10. Tao-Cheng, J., Vinade, L., Winters, C., Reese, T. and Dosemeci, A. Inhibition of phosphatase activity facilitates the formation and maintenance of NMDA-induced calcium/calmodulin-dependent protein kinase II clusters in hippocampal neurons. Neuroscience 130 (2005) 651–656.

    Article  PubMed  CAS  Google Scholar 

  11. Ashpole, N., Song, W., Brustovetsky, T., Engleman, E., Brustovetsky, N., Cummins, T. and Hudmon, A. Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibition induces neurotoxicity via dysregulation of glutamate/calcium signaling and hyperexcitability. J. Biol. Chem. 287 (2012) 8495–8506.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. Shim, S., Wang, J., Asada, N., Neumayer, G., Tran, H., Ishiguro, K., Sanada, K., Nakatani, Y. and Nguyen, M. Protein 600 is a microtubule/endoplasmic reticulum-associated protein in CNS neurons. J. Neurosci. 28 (2008) 3604–3614.

    Article  PubMed  CAS  Google Scholar 

  13. Tasaki, T., Kim, S., Zakrzewska, A., Lee, B., Kang, M., Yoo, Y., Cha-Molstad, H., Hwang, J., Soung, N., Sung, K., Kim, S.-H., Nguyen, M., Sun, M., Yi, E., Kim, B. and Kwon, Y. UBR box N-recognin-4 (UBR4), an N-recognin of the N-end rule pathway, and its role in yolk sac vascular development and autophagy. Proc. Natl. Acad. Sci. U.S.A. 4 (2013) 3800–3805.

    Article  Google Scholar 

  14. Nakatani, Y., Konishi, H., Vassilev, A., Kurooka, H., Ishiguro, K., Sawada, J., Ikura, T., Korsmeyer, S. J., Qin, J. and Herlitz, A.M. p600, a unique protein required for membrane morphogenesis and cell survival. Proc. Natl. Acad. Sci. U.S.A. 102 (2005) 15093–15098.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  15. Tasaki, T., Mulder, L., Iwamatsu, A., Lee, M., Davydov, I., Varshavsky, A., Muesing, M. and Kwon, Y. A family of mammalian E3 ubiquitin ligases that contain the UBR box motif and recognize N-degrons. Mol. Cell. Biol. 25 (2005) 7120–7136.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. DeMasi, J. and Huh, K. Bovine papillomavirus E7 transformation function correlates with cellular p600 protein binding. Proc. Natl. Acad. Sci. U.S.A. 103 (2005) 11486–11491.

    Article  Google Scholar 

  17. Belzil, C., Neumayer, G., Vassilev, A.P., Yap, K.L., Konishi, H., Rivest, S., Sanada, K., Ikura, M., Nakatani, Y. and Nguyen, M.D. A Ca2+-dependent mechanism of neuronal survival mediated by the microtubule-associated protein p600. J. Biol. Chem. 288 (2013) 24452–24464.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  18. Rose, J., Jin, S. and Craig, A. Heterosynaptic molecular dynamics: Locally induced propagating synaptic accumulation of CaM kinase II. Neuron 61 (2009) 351–358.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Colicos, M., Collins, B., Sailor, M. and Goda, Y. Remodeling of synaptic actin induced by photoconductive stimulation. Cell. 107 (2001) 605–616.

    Article  PubMed  CAS  Google Scholar 

  20. Goda, Y. and Colicos, M. Photoconductive stimulation of neurons cultured on silicon wafers. Nat. Prot. 1 (2006) 461–467.

    Article  CAS  Google Scholar 

  21. Lemieux, M., Labrecque, S., Tardif, C., Labrie-Dion, E., Lebel, E. and De Koninck, P. Translocation of CaMKII to dendritic microtubules supports the plasticity of local synapses. J. Biol. Chem. 198 (2012) 1055–1073.

    CAS  Google Scholar 

  22. Furukawa, K. and Mattson, M. Taxol stabilizes [Ca2+]i and protects hippocampal neurons against excitotoxicity. Brain Res. 689 (1995) 141–146.

    Article  PubMed  CAS  Google Scholar 

  23. Siman, R. and Noszek, J. Excitatory amino acids activate calpain I and induce structural protein breakdown in vivo. Neuron 1 (1988) 279–287.

    Article  PubMed  CAS  Google Scholar 

  24. Pettigrew, L., Holtz, M., Craddock, S., Minger, L., Hall, N. and Geddes, J. Microtubular proteolysis in focal cerebral ischemia. J. Cereb. Blood Flow Metab. 16 (1996) 1189–1202.

    Article  PubMed  CAS  Google Scholar 

  25. Chang, B., Mukherji, S. and Soderling, T. Calcium/calmodulin-dependent protein kinase II inhibitor protein: localization of isoforms in rat brain. Neuroscience 102 (2001) 767–777.

    Article  PubMed  CAS  Google Scholar 

  26. Hudmon, A., Kim, S., Kolb, S., Stoops, J. and Waxham, M. Light scattering and transmission electron microscopy studies reveal a mechanism for calcium/calmodulin-dependent protein kinase II self-association. J. Neurochem. 76 (2001) 1364–1375.

    Article  PubMed  CAS  Google Scholar 

  27. Barria, A., Muller, D., Derkach, V., Griffith, L. and Soderling, T. Regulatory phosphorylation of AMPA-type glutamate receptors by CaM-KII during long-term potentiation. Science 276 (1997) 2042–2045.

    Article  PubMed  CAS  Google Scholar 

  28. Barria, A., Derkach, V. and Soderling, T. Identification of the Ca2+/calmodulindependent protein kinase II regulatory phosphorylation site in the α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate-type glutamate receptor. J. Biol. Chem. 272 (1997) 32727–32730.

    Article  PubMed  CAS  Google Scholar 

  29. Hudmon, A., Schulman, H., Kim, J., Maltez, J., Tsien, R. and Pitt, G. CaMKII tethers to L-type Ca2+ channels, establishing a local and dedicated integrator of Ca2+ signals for facilitation. J. Cell. Biol. 171 (2005) 537–547.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  30. Sanhueza, M., Fernandez-Villalobos, G., Stein, I.S., Kasumova, G., Zhang, P., Bayer, K.U., Otmakhov, N., Hell, J.W. and Lisman, J. Role of the CaMKII/NMDA receptor complex in the maintenance of synaptic strength. J. Neurosci. 31 (2011) 9170–9178.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  31. Vallano, M.L., Goldenring, J.R., Lasher, R.S. and Delorenzo, R.J. Association of calcium/calmodulin-dependent kinase with cytoskeletal preparations: phosphorylation of tubulin, neurofilament, and microtubuleassociated proteins proteins. Ann. N.Y. Acad. Sci. 466 (1986) 357–374.

    Article  PubMed  CAS  Google Scholar 

  32. Wandosell, F., Serrano, L., Hernández, M.A. and Avila, J. Phosphorylation of tubulin by a calmodulin-dependent protein kinase. J. Biol. Chem. 261 (1986) 10332–10339.

    PubMed  CAS  Google Scholar 

  33. Craddock, T.J., Tuszynski, J.A. and Hameroff, S. Cytoskeletal signaling: is memory encoded in microtubule lattices by CaMKII phosphorylation? PLoS Comp. Biol. 8 (2012) e1002421.

    Article  CAS  Google Scholar 

  34. Goldschmidt, R. and Steward, O. Preferential neurotoxicity of colchicine for granule cells of the dentate. Proc. Natl. Acad. Sci. USA 77 (1980) 3047–3051.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  35. Kristensen, B., Noer, H., Gramsbergen, J., Zimmer, J. and Noraberg, J. Colchicine induces apoptosis in organotypic hippocampal slice cultures. Brain Res. 964 (2003) 264–278.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Hotchkiss Brain Institute, University of Calgary, Departments of Clinical Neurosciences, Cell Biology & Anatomy, Biochemistry & Molecular Biology, 3330 Hospital Drive NW, Calgary, Alberta, Canada, T2N 4N1

    Camille Belzil, Tim Ramos, Michael A. Colicos & Minh Dang Nguyen

  2. Molecular Genetics Research Laboratory, Graduate School of Science, University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo, 113-0033, Japan

    Kamon Sanada

Authors
  1. Camille Belzil
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tim Ramos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kamon Sanada
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael A. Colicos
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Minh Dang Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Minh Dang Nguyen.

Electronic supplementary material

Supplementary material, approximately 283 KB.

Supplementary material, approximately 426 KB.

Supplementary material, approximately 2.83 MB.

Supplementary material, approximately 1.24 MB.

Supplementary material, approximately 118 KB.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Belzil, C., Ramos, T., Sanada, K. et al. p600 stabilizes microtubules to prevent the aggregation of CaMKIIα during photoconductive stimulation. Cell Mol Biol Lett 19, 381–392 (2014). https://doi.org/10.2478/s11658-014-0201-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-014-0201-9

Keywords

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us