The physiological state of the cell is controlled by signal transduction mechanisms which regulate the balance between protein kinase and protein phosphatase activities1. Here we report that a single protein can, depending on which particular amino-acid residue is phosphorylated, function either as a kinase or phosphatase inhibitor. DARPP-32 (dopamine and cyclic AMP-regulated phospho-protein, relative molecular mass 32,000) is converted into an inhibitor of protein phosphatase 1 when it is phosphorylated by protein kinase A (PKA) at threonine 34 (refs 2, 3). We find that DARPP-32 is converted into an inhibitor of PKA when phosphorylated at threonine 75 by cyclin-dependent kinase 5 (Cdk5). Cdk5 phosphorylates DARPP-32 in vitro and in intact brain cells. Phospho-Thr 75 DARPP-32 inhibits PKA in vitro by a competitive mechanism. Decreasing phospho-Thr 75 DARPP-32 in striatal slices, either by a Cdk5-specific inhibitor or by using genetically altered mice, results in increased dopamine-induced phosphorylation of PKA substrates and augmented peak voltage-gated calcium currents. Thus DARPP-32 is a bifunctional signal transduction molecule which, by distinct mechanisms, controls a serine/threonine kinase and a serine/threonine phosphatase.
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Hunter,T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 80, 225–236 (1995).
Hemmings,H. C. Jr, Greengard,P., Tung,H. Y. L. & Cohen,P. DARPP-32, a dopamine-regulated neuronal phosphoprotein, is a potent inhibitor of protein phosphatase-1. Nature 310, 503–505 (1984).
Greengard,P., Allen,P. B. & Nairn,A. C. Beyond the dopamine receptor: the DARPP-32/protein phosphatase-1 cascade. Neuron 23, 435–447 (1999).
Fienberg,A. A. et al. DARPP-32: Regulator of the efficacy of dopaminergic neurotransmission. Science 281, 838–842 (1998).
Lew,J. et al. A brain-specific activator of cyclin-dependent kinase 5. Nature 371, 423–426 (1994).
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).
Meijer,L. et al. Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. Eur. J. Biochem. 243, 527–536 (1997).
Ouimet,C. C., Langley-Gullion,K. C. & Greengard,P. Quantitative immunocytochemistry of DARPP-32-expressing neurons in the rat caudatoputamen. Brain Res. 808, 8–12 (1998).
Hemmings,H. C. Jr, Girault,J.-A., Williams,K. R., LoPresti,M. B. & Greengard,P. ARPP-21, a cyclic AMP-regulated phosphoprotein (Mr = 21,000) enriched in dopamine-innervated brain regions: Amino-acid sequence of the site phosphorylated by cyclic AMP in intact cells and kinetic studies of its phosphorylation in vitro. J. Biol. Chem. 264, 7726–7733 (1989).
Roche,K. W., O'Brien,R. J., Mammen,A. L., Bernhardt,J. & Huganir,R. L. Characterization of multiple phosphorylation sites on the AMPA receptor GluRI subunit. Neuron 16, 1179–1188 (1996).
Horiuchi,A., Williams,K. R., Kurihara,T., Nairn,A. C. & Greengard,P. Purification and cDNA cloning of ARPP-16, a cAMP-regulated phosphoprotein enriched in basal ganglia, and of a related phosphoprotein, ARPP-19. J. Biol. Chem. 265, 9476–9484 (1990).
Surmeier,D. J., Bargas,J. Hemmings,H. C. Jr, Nairn,A. C. & Greengard,P. Modulation of calcium currents by a D1 dopaminergic protein kinase/phosphatase cascade in rat neostriatal neurons. Neuron 14, 385–397 (1995).
Gray,P. C., Scott,J. D. & Caterall,W. A. Regulation of ion channels by cAMP-dependent protein kinase and A-kinase anchoring proteins. Curr. Opin. Neurobiol. 8, 330–334 (1998).
Nicola,S. M., Surmeier,J. D. & Malenka,R. C. Dopaminergic modulation of neuronal excitability in the striatum and nucleus accumbens. Annu. Rev. Neurosci. (in the press).
Pawson,T. & Scott,J. D. Signaling through scaffold, anchoring, and adaptor proteins. Science 278, 2075–2080 (1997).
Klauck,T. M. et al. Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science 271, 1589–1592 (1996).
Westphal,R. S., Anderson, K. A., Means,A. R. & Wadzinski,B. E. A signaling complex of Ca2+-calmodulin-dependent protein kinase IV and protein phosphatase 2A. Science 280, 1258–1261 (1998).
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).
Czernik,A. J., Mathers,J., Tsou,K., Greengard,P. & Mische,S. M. in Regulatory Protein Modification, Neuromethods vol. 30 (ed. Hemmings, H. C. Jr) 219–246 (Humana, Totowa, New Jersey, 1997).
Girault,J.-A., Hemmings,H. C. Jr, Williams,K. R., Nairn,A. C. & Greengard,P. Phosphorylation of DARPP-32, a dopamine- and cAMP-regulated phosphoprotein, by casein kinase II. J. Biol. Chem. 264, 21748–21759 (1989).
Nishi,A., Snyder,G. L. & Greengard,P. Bidirectional regulation of DARPP-32 phosphorylation by dopamine. J. Neurosci. 17, 8147–8155 (1997).
Snyder,G. L. et al. Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: Regulation by factors other than dopamine. J. Neurosci. 12, 3071–3083 (1992).
Caporaso,G. et al. Dopamine and drugs of abuse modulate phosphorylation of ARPP-21, a cyclic AMP-regulated phosphoprotein enriched in the neostriatum. Neuropharmacology (in press).
We thank A. Horiuchi for construction of the Thr75A mutant plasmid; S. Rakhilin for recombinant ARPP-21; I. Dulubova for phospho-Ser88 ARPP-16 antibody; J. Wang for recombinant Cdk5/p25; J. Volker for help in providing p35-/- mice; and the Rockefeller University Protein/DNA Technology Center. This work was supported by a National Research Service Award (J.B.) and funding from the National Institute of Mental Health and the National Institute of Drug Abuse (P.G.).
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Bibb, J., Snyder, G., Nishi, A. et al. Phosphorylation of DARPP-32 by Cdk5 modulates dopamine signalling in neurons. Nature 402, 669–671 (1999) doi:10.1038/45251
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