Translocation of calmodulin to the nucleus supports CREB phosphorylation in hippocampal neurons

Abstract

Activation of the transcription factor CREB is thought to be important in the formation of long-term memory in several animal species1,2,3. The phosphorylation of a serine residue at position 133 of CREB is critical for activation of CREB4. This phosphorylation is rapid when driven by brief synaptic activity in hippocampal neurons5. It is initiated by a highly local, rise in calcium ion concentration5 near the cell membrane, but culminates in the activation of a specific calmodulin-dependent kinase known as CaMK IV (ref. 7), which is constitutively present in the neuronal nucleus7,8. It is unclear how the signal is conveyed from the synapse to the nucleus. We show here that brief bursts of activity cause a swift (1 min) translocation of calmodulin from the cytoplasm to the nucleus, and that this translocation is important for the rapid phosphorylation of CREB. Certain Ca2+ entry systems (L-type Ca2+ channels and NMDA receptors) are able to cause mobilization of calmodulin, whereas others (N- and P/Q-type Ca2+ channels) are not. This translocation of calmodulin provides a form of cellular communication that combines the specificity of local Ca2+ signalling with the ability to produce action at a distance.

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Figure 1: Rapid activity-dependent translocation of calmodulin to the nucleus in hippocampal CA3/CA1 pyramidal neurons.
Figure 2: Selective control of activity-dependent CaM translocation by NMDA receptors and L-type Ca2+ channels.
Figure 3: Close relationship between activity-dependent CaM translocation and CREB phosphorylation.
Figure 4: Requirement for nuclear CaM translocation in activity-dependent CREBphosphorylation.
Figure 5: Causal link between nuclear CaM and activity-dependent CREB phosphorylation in hippocampal neurons.

References

  1. 1

    Dash, P. K., Hochner, B. & Kandel, E. R. Injection of the cAMP-responsive element into the nucleus of Aplysia sensory neurons blocks long-term facilitation. Nature 345, 718–721 (1990).

  2. 2

    Bourtchuladze, R. et al. Deficient long-term memory in mice with a targeted mutation of the cAMP-responsive element-binding protein. Cell 79, 59–68 (1994).

  3. 3

    Yin, J. C. et al. Induction of a dominant negative CREB transgene specifically blocks long-term memory in Drosophila. Cell 79, 49–58 (1994).

  4. 4

    Gonzalez, G. A. & Montminy, M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell 59, 675–680 (1989).

  5. 5

    Deisscroth, K., Bito, H. & Tsien, R. W. Signaling from synapse to nucleus: postsynaptic CREB phosphorylation during multiple forms of hippocampal synaptic plasticity. Neuron 16, 89–101 (1996).

  6. 6

    Kavalali, E., Zhuo, M., Bito, H. & Tsien, R. W. Dendritic Ca2+ channels characterized by recordings from isolated hippocampal dendritic segments. Neuron 18, 651–663 (1997).

  7. 7

    Bito, H., Deisseroth, K. & Tsien, R. W. CREB phosphorylation and dephosphorylation: a Ca2+- and stimulus duration-dependent switch for hippocampal gene expression. Cell 87, 1203–1214 (1996).

  8. 8

    Nakamura, Y., Okuno, S., Sato, F. & Fujisawa, H. An immunohistochemical study of Ca2+/calmodulin-dependent protein kinase IV in the rat central nervous system: light and electron microscopic observations. Neuroscience 68, 181–194 (1995).

  9. 9

    Sheng, M., Thompson, M. A. & Greenberg, M. E. CREB: a Ca2+-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science 252, 1427–1430 (1991).

  10. 10

    Dash, P. K., Karl, K. A., Colicos, M. A., Prywes, R. & Kandel, E. R. cAMP response element-binding protein is activated by Ca2+/calmodulin- as well as cAMP-dependent protein kinase. Proc. Natl Acad. Sci. USA 88, 5061–5065 (1991).

  11. 11

    Luby-Phelps, K., Hori, M., Phelps, J. M. & Won, D. Ca2+-regulated dynamic compartmentalization of calmodulin in living smooth muscle cells. J. Biol. Chem. 270, 21532–21538 (1995).

  12. 12

    Pruschy, M., Ju, Y., Spitz, L., Carafoli, E. & Goldfarb, D. S. Facilitated nuclear transport of calmodulin in tissue culture cells. J. Cell Biol. 127, 1527–1536 (1994).

  13. 13

    Vendrell, M., Pujol, M. J., Tusell, J. M. & Serratosa, J. Effect of different convulsants on calmodulin levels and proto-oncogene c-fos expression in the central nervous system. Brain Res. Mol. Brain Res. 14, 285–292 (1992).

  14. 14

    Mitsui, K., Brady, M., Palfrey, H. C. Nairn, A. C. Purification and characterization of calmodulin-dependent protein kinase III from rabbit reticulocytes and rat pancreas. J. Biol. Chem. 268, 13422–13433 (1993).

  15. 15

    Hahn, K. W., Waggoner, A. S. & Taylor, D. L. Acalcium-sensitive fluorescent analog of calmodulin based on a novel calmodulin-binding fluorophore. J. Biol. Chem. 265, 20335–20345 (1990).

  16. 16

    Westenbroek, R. E., Ahlijanian, M. K. & Catterall, W. A. Clustering of L-type Ca2+ channels at the base of major dendrites in hippocampal pyramidal neurons. Nature 347, 281–284 (1990).

  17. 17

    Magee, J. C. & Johnston, D. Characterization of single voltage-gated Na2+ and Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons. J. Physiol. 487, 67–90 (1995).

  18. 18

    Morgan, J. I. & Curran, T. Role of ion flux in the control of c-fos expression. Nature 322, 552–555 (1986).

  19. 19

    Murphy, T. H., Worley, P. F. & Barahan, J. M. L-type voltage-sensitive calcium channels mediate synaptic activation of immediate early genes. Neuron 7, 625–635 (1991).

  20. 20

    Ginty, D. D. et al. Regulation of CREB phosphorylation in the suprachiasmatic nucleus by light and a circadian clock. Science 260, 238–241 (1993).

  21. 21

    Hagiwara, M. et al. Coupling of hormonal stimulation and transcription via the cyclic AMP responsive factor CREB is rate limited by nuclear entry of protein kinase A. Mol. Cell. Biol. 13, 4852–4859 (1993).

  22. 22

    Harootunian, A. T. et al. Movement of the free catalytic subunit of cAMP-dependent protein kinase into and out of the nucleus can be explained by diffusion. Mol. Biol. Cell. 4, 993–1002 (1993).

  23. 23

    Xia, Z., Dudek, H., Miranti, C. K. & Greenberg, M. E. Calcium influx via the NMDA receptor induces immediate early gene transcription by a MAP kinase/ERK-dependent mechanism. J. Neurosci. 16, 5425–5436 (1996).

  24. 24

    Wang, J., Campos, B., Jamieson, G. A. J, Kaetzel, M. A. & Dedman, J. R. Functional elimination of calmodulin within the nucleus by targeted expression of an inhibitor peptide. J. Biol. Chem. 270, 30245–30248 (1995).

  25. 25

    Hardingham, G. E., Chawla, S., Johnson, C. M. & Bading, H. Distinct functions of nuclear and cytoplasmic calcium in the control of gene expression. Nature 385, 260–265 (1997).

  26. 26

    Means, A. R. et al. Anovel Ca2+/calmodulin-dependent protein kinase and a male germ cell-specific calmodulin-binding protein are derived from the same gene. Mol. Cell. Biol. 11, 3960–3971 (1991).

  27. 27

    Bito, H., Deisseroth, K. & Tsien, R. W. Ca2+-dependent regulation in neuronal gene expression. Curr. Opin. Neurobiol. 7, 419–429 (1997).

  28. 28

    Tokumitsu, H. & Soderling, T. R. Requirements for calcium and calmodulin in the calmodulin kinase activation cascade. J. Biol. Chem. 271, 5617–5622 (1996).

  29. 29

    Sacks, D. B., Porter, S. E., Ladenson, J. H. & McDonald, J. M. Monoclonal antibody to calmodulin: development, characterization, and comparison with polyclonal anti-calmodulin antibodies. Analyt. Biochem. 194, 369–377 (1991).

  30. 30

    Hulen, D., Baron, A., Salisbury, J. & Clarke, M. Production and specificity of monoclonal antibodies against calmodulin from Dicytostelium discoideum. Cell. Motil. Cytoskel. 18, 113–122 (1991).

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Acknowledgements

We thank A. Nairn for antibodies; J. Dedman, J. Wang and M. Kaetzel for nCaMBP reagents; D. L. Taylor and J. Montibeller for mCaM; H. Schulman and H. Bito for advice and for comments on the manuscript; and R. Lewis, T. Schwarz and L. Stryer for comments on the manuscript. Supported by grants from the Silvio Conte-NIMH Center for Neuroscience Research, the McKnight Foundation, the Mathers Charitable Trust, and the NIH Medical Scientist Training Program.

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Correspondence to Richard W. Tsien.

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