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Phosphorylation of WAVE1 regulates actin polymerization and dendritic spine morphology

Abstract

WAVE1—the Wiskott–Aldrich syndrome protein (WASP)-family verprolin homologous protein 1—is a key regulator of actin-dependent morphological processes1 in mammals, through its ability to activate the actin-related protein (Arp2/3) complex. Here we show that WAVE1 is phosphorylated at multiple sites by cyclin-dependent kinase 5 (Cdk5) both in vitro and in intact mouse neurons. Phosphorylation of WAVE1 by Cdk5 inhibits its ability to regulate Arp2/3 complex-dependent actin polymerization. Loss of WAVE1 function in vivo or in cultured neurons results in a decrease in mature dendritic spines. Expression of a dephosphorylation-mimic mutant of WAVE1 reverses this loss of WAVE1 function in spine morphology, but expression of a phosphorylation-mimic mutant does not. Cyclic AMP (cAMP) signalling reduces phosphorylation of the Cdk5 sites in WAVE1, and increases spine density in a WAVE1-dependent manner. Our data suggest that phosphorylation/dephosphorylation of WAVE1 in neurons has an important role in the formation of the filamentous actin cytoskeleton, and thus in the regulation of dendritic spine morphology.

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Figure 1: WAVE1 interacts with, and is phosphorylated by, P35/Cdk5.
Figure 2: Inhibition of Arp2/3-complex-mediated actin polymerization by phosphorylation of WAVE1.
Figure 3: Role of WAVE1 in spine morphogenesis.
Figure 4: cAMP-dependent dephosphorylation of WAVE1 and its role in spine formation.

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Acknowledgements

We thank W.-B. Gan for assisting us with the DiI staining method; L.-H. Tsai for providing P35 cDNA; A. Yamamoto for Nap1 cDNA; and P. Aspenström for N-WASP antibody. We also thank T. D. Pollard for suggestions. This work was supported by the postdoctoral fellowship programme of the Korea Science & Engineering Foundation (KOSEF) (to J.Y.S.), and funding from the F.M. Kirby foundation (to P.G.), the Picower Foundation (to P.G.), the National Institute of Mental Health, the National Institute of Drug Abuse and the National Institute on Aging (to A.C.N. and P.G.). Author Contributions Y.K., J.Y.S., I.C. and K.-W.L. performed experiments; J.M.H. and A.M.K. assisted in experiments; J.-H.A., S.P.K., A.S., B.B. and J.D.S. provided reagents; J.B.P. and S.H.R. performed MALDI-TOF mass spectrometry; Y.K., A.C.N. and P.G. designed experiments and wrote the manuscript.

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Correspondence to Paul Greengard.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

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Supplementary Notes

This file contains Supplementary Figures and Legends 1–7, Supplementary Table 1 and Supplementary Methods. The Supplementary Figures show the interaction of P35 with the WAVE complex, phosphorylation of WAVE1, actin polymerization catalyzed by WAVE1 and Cdk5 activity in striatal slices following forskolin treatment. The Supplementary Table provides stoichiometry of phosphorylation of WAVE1. The Supplementary Methods provide technical details of experimental methods used in this study. (PDF 622 kb)

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Kim, Y., Sung, J., Ceglia, I. et al. Phosphorylation of WAVE1 regulates actin polymerization and dendritic spine morphology. Nature 442, 814–817 (2006). https://doi.org/10.1038/nature04976

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