Staurosporine inhibits agrin-induced acetylcholine receptor phosphorylation and aggregation

doi:10.1083/jcb.125.3.661

This Article

	Full Text (PDF, 1074K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Wallace, B. G.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Wallace, B. G.


Social Bookmarking

	What's this?

ARTICLES

Staurosporine inhibits agrin-induced acetylcholine receptor phosphorylation and aggregation

BG Wallace
Department of Physiology, University of Colorado Health Sciences Center, Denver 80262.

Agrin, a protein that mediates nerve-induced acetylcholine receptor (AChR) aggregation at developing neuromuscular junctions, has been shown to cause an increase in phosphorylation of the beta, gamma, and delta subunits of AChRs in cultured myotubes. As a step toward understanding the mechanism of agrin-induced AChR aggregation, we examined the effects of inhibitors of protein kinases on AChR aggregation and phosphorylation in chick myotubes in culture. Staurosporine, an antagonist of both protein serine and tyrosine kinases, blocked agrin-induced AChR aggregation in a dose-dependent manner; 50% inhibition occurred at approximately 2 nM. The extent of inhibition was independent of agrin concentration, suggesting an effect downstream of the interaction of agrin with its receptor. Staurosporine blocked agrin-induced phosphorylation of the AChR beta subunit, which occurs at least in part on tyrosine residues, but did not reduce phosphorylation of the gamma and delta subunits, which occurs on serine/threonine residues. Staurosporine also prevented the agrin- induced decrease in the rate at which AChRs are extracted from intact myotubes by mild detergents. H-7, an antagonist of protein serine kinases, inhibited agrin-induced phosphorylation of the gamma and delta subunits but did not block agrin-induced phosphorylation of the AChR beta subunit, AChR aggregation, or the decrease in AChR extractability. The results provide support for the hypothesis that tyrosine phosphorylation of the beta subunit plays a role in agrin-induced AChR aggregation.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Bruneau, E., Akaaboune, M. (2007). The Dynamics of the Rapsyn Scaffolding Protein at Individual Acetylcholine Receptor Clusters. J. Biol. Chem. 282: 9932-9940 [Abstract] [Full Text]
Bruneau, E. G., Akaaboune, M. (2006). The dynamics of recycled acetylcholine receptors at the neuromuscular junction in vivo. Development 133: 4485-4493 [Abstract] [Full Text]
Jacobson, C., Duggan, D., Fischbach, G. (2004). Neuregulin induces the expression of transcription factors and myosin heavy chains typical of muscle spindles in cultured human muscle. Proc. Natl. Acad. Sci. USA 101: 12218-12223 [Abstract] [Full Text]
Marangi, P. A., Wieland, S. T., Fuhrer, C. (2002). Laminin-1 redistributes postsynaptic proteins and requires rapsyn, tyrosine phosphorylation, and Src and Fyn to stably cluster acetylcholine receptors. JCB 157: 883-895 [Abstract] [Full Text]
Mohamed, A. S., Rivas-Plata, K. A., Kraas, J. R., Saleh, S. M., Swope, S. L. (2001). Src-Class Kinases Act within the Agrin/MuSK Pathway to Regulate Acetylcholine Receptor Phosphorylation, Cytoskeletal Anchoring, and Clustering. J. Neurosci. 21: 3806-3818 [Abstract] [Full Text]
Smith, C. L., Mittaud, P., Prescott, E. D., Fuhrer, C., Burden, S. J. (2001). Src, Fyn, and Yes Are Not Required for Neuromuscular Synapse Formation But Are Necessary for Stabilization of Agrin-Induced Clusters of Acetylcholine Receptors. J. Neurosci. 21: 3151-3160 [Abstract] [Full Text]
Borges, L. S., Ferns, M. (2001). Agrin-Induced Phosphorylation of the Acetylcholine Receptor Regulates Cytoskeletal Anchoring and Clustering. JCB 153: 1-12 [Abstract] [Full Text]
Bose, C. M., Qiu, D., Bergamaschi, A., Gravante, B., Bossi, M., Villa, A., Rupp, F., Malgaroli, A. (2000). Agrin Controls Synaptic Differentiation in Hippocampal Neurons. J. Neurosci. 20: 9086-9095 [Abstract] [Full Text]
NGHIÊM, H.-O., BETTENDORFF, L., CHANGEUX, J.-P. (2000). Specific phosphorylation of Torpedo 43K rapsyn by endogenous kinase(s) with thiamine triphosphate as the phosphate donor. FASEB J. 14: 543-554 [Abstract] [Full Text]
Marchand, S., Bignami, F., Stetzkowski-Marden, F., Cartaud, J. (2000). The Myristoylated Protein Rapsyn is Cotargeted with the Nicotinic Acetylcholine Receptor to the Postsynaptic Membrane via the Exocytic Pathway. J. Neurosci. 20: 521-528 [Abstract] [Full Text]
Hilgenberg, L. G. W., Hoover, C. L., Smith, M. A. (1999). Evidence of an Agrin Receptor in Cortical Neurons. J. Neurosci. 19: 7384-7393 [Abstract] [Full Text]
Mohamed, A. S., Swope, S. L. (1999). Phosphorylation and Cytoskeletal Anchoring of the Acetylcholine Receptor by Src Class Protein-tyrosine Kinases. ACTIVATION BY RAPSYN. J. Biol. Chem. 274: 20529-20539 [Abstract] [Full Text]
Cornish, T, Chi, J, Johnson, S, Lu, Y, Campanelli, J. (1999). Globular domains of agrin are functional units that collaborate to induce acetylcholine receptor clustering. J. Cell Sci. 112: 1213-1223 [Abstract]
Hopf, C., Hoch, W. (1998). Dimerization of the Muscle-specific Kinase Induces Tyrosine Phosphorylation of Acetylcholine Receptors and Their Aggregation on the Surface of Myotubes. J. Biol. Chem. 273: 6467-6473 [Abstract] [Full Text]
Burden, S. J. (1998). The formation of neuromuscular�synapses. Genes Dev. 12: 133-148 [Full Text]
Sugiyama, J.E., Glass, D.J., Yancopoulos, G.D., Hall, Z.W. (1997). Laminin-induced Acetylcholine Receptor Clustering: An Alternative Pathway. JCB 139: 181-191 [Abstract] [Full Text]
Glass, D. J., Apel, E. D., Shah, S., Bowen, D. C., DeChiara, T. M., Stitt, T. N., Sanes, J. R., Yancopoulos, G. D. (1997). Kinase domain of the muscle-specific receptor tyrosine kinase (MuSK) is sufficient for phosphorylation but not clustering of acetylcholine receptors: Required role for the MuSK�ectodomain?. Proc. Natl. Acad. Sci. USA 94: 8848-8853 [Abstract] [Full Text]
Colledge, M., Froehner, S. C. (1997). Tyrosine Phosphorylation of Nicotinic Acetylcholine Receptor Mediates Grb2 Binding. J. Neurosci. 17: 5038-5045 [Abstract] [Full Text]
Cohen, M. W., Jacobson, C., Yurchenco, P. D., Morris, G. E., Carbonetto, S. (1997). Laminin-induced Clustering of Dystroglycan on Embryonic Muscle Cells: Comparison with Agrin-induced Clustering. JCB 136: 1047-1058 [Abstract] [Full Text]
Fuhrer, C., Hall, Z. W. (1996). Functional Interaction of Src Family Kinases with the Acetylcholine Receptor in C2 Myotubes. J. Biol. Chem. 271: 32474-32481 [Abstract] [Full Text]
Bowen, D. C., Sugiyama, J., Ferns, M., Hall, Z. W. (1996). Neural Agrin Activates a High-Affinity Receptor in C2 Muscle Cells that Is Unresponsive to Muscle Agrin. J. Neurosci. 16: 3791-3797 [Abstract] [Full Text]
Kirsch, J., Kroger, S. (1996). {blacksquare} REVIEW : Postsynaptic Anchoring of Receptors: A Cellular Approach to Neuronal and Muscular Sensitivity. Neuroscientist 2: 100-108 [Abstract]
Brisseau, G. F., Grinstein, S., Hackam, D. J., Nordstr�m, T., Manolson, M. F., Khine, A. A., Rotstein, O. D. (1996). Interleukin-1 Increases Vacuolar-type H[IMAGE]-ATPase Activity in Murine Peritoneal Macrophages. J. Biol. Chem. 271: 2005-2011 [Abstract] [Full Text]
Escher, G, Bechade, C, Levi, S, Triller, A (1996). Axonal targeting of agrin in cultured rat dorsal horn neurons. J. Cell Sci. 109: 2959-2966 [Abstract]
Cartaud, A, Ludosky, M., Haasemann, M, Jung, D, Campbell, K, Cartaud, J (1996). Non-neural agrin codistributes with acetylcholine receptors during early differentiation of Torpedo electrocytes. J. Cell Sci. 109: 1837-1846 [Abstract]
Campanelli, J., Gayer, G., Scheller, R. (1996). Alternative RNA splicing that determines agrin activity regulates binding to heparin and alpha-dystroglycan. Development 122: 1663-1672 [Abstract]
Wang, Z.-Z., Fuhrer, C., Shtrom, S., Sugiyama, J.E., Ferns, M.J., Hall, Z.W. (1996). The Nicotinic Acetylcholine Receptor at the Neuromuscular Junction: Assembly and Tyrosine Phosphorylation. Cold Spring Harb Symp Quant Biol 61: 363-371 [Abstract]
Glass, D.J., DeChiara, T.M., Stitt, T.N., DiStefano, P.S., Valenzuela, D.M., Yancopoulos, G.D. (1996). The Receptor Tyrosine Kinase MuSK Is Required for Neuromuscular Junction Formation and Is a Functional Receptor for Agrin. Cold Spring Harb Symp Quant Biol 61: 435-444 [Abstract]
Lau, L.-F., Huganir, R. L. (1995). Differential Tyrosine Phosphorylation of N-Methyl-D-aspartate Receptor Subunits. J. Biol. Chem. 270: 20036-20041 [Abstract] [Full Text]
Mittaud, P., Marangi, P. A., Erb-Vogtli, S., Fuhrer, C. (2001). Agrin-induced Activation of Acetylcholine Receptor-bound Src Family Kinases Requires Rapsyn and Correlates with Acetylcholine Receptor Clustering. J. Biol. Chem. 276: 14505-14513 [Abstract] [Full Text]