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J Biol Chem, Vol. 273, Issue 18, 11321-11326, May 1, 1998

Evidence for in Situ and in Vitro Association between -Dystroglycan and the Subsynaptic 43K Rapsyn Protein
CONSEQUENCE FOR ACETYLCHOLINE RECEPTOR CLUSTERING AT THE SYNAPSE

Annie Cartaud, Sébastien Coutant, Tamara C. Petrucci^§, and Jean Cartaud

From  Biologie Cellulaire des Membranes, Département de Biologie Supramoléculaire et Cellulaire, Institut Jacques Monod, UMR 9922, CNRS et Université Paris VII, 2 Place Jussieu, 75251 Paris Cédex 05, France and ^§ Biologia Cellulare, Istituto Superiore di Sanità, V. le Regina Elena 299, Roma 00161, Italy

The accumulation of dystrophin and associated proteins at the postsynaptic membrane of the neuromuscular junction and their co-distribution with nicotinic acetylcholine receptor (AChR) clusters in vitro suggested a role for the dystrophin complex in synaptogenesis. Co-transfection experiments in which - and -dystroglycan form a complex with AChR and rapsyn, a peripheral protein required for AChR clustering (Apel, D. A., Roberds, S. L., Campbell, K. P., and Merlie, J. P. (1995) Neuron 15, 115-126), suggested that rapsyn functions as a link between AChR and the dystrophin complex. We have investigated the interaction between rapsyn and -dystroglycan in Torpedo AChR-rich membranes using in situ and in vitro approaches. Cross-linking experiments were carried out to study the topography of postsynaptic membrane polypeptides. A cross-linked product of 90 kDa was labeled by antibodies to rapsyn and -dystroglycan; this demonstrates that these polypeptides are in close proximity to one another. Affinity chromatography experiments and ligand blot assays using rapsyn solubilized from Torpedo AChR-rich membranes and constructs containing -dystroglycan C-terminal fragments show that a rapsyn-binding site is present in the juxtamembranous region of the cytoplasmic tail of -dystroglycan. These data point out that rapsyn and dystroglycan interact in the postsynaptic membrane and thus reinforce the notion that dystroglycan could be involved in synaptogenesis.

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