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Regulation of capacitative calcium entries by 1-syntrophin: association of TRPC1 with dystrophin complex and the PDZ domain of 1-syntrophin

Aurélie Vandebrouck^*, Jessica Sabourin^*, Jérôme Rivet, Haouria Balghi^*, Stéphane Sebille^*, Alain Kitzis^*,, Guy Raymond^*, Christian Cognard^*, Nicolas Bourmeyster^*, and Bruno Constantin^*,1

^* Institut de Physiologie et Biologie Cellulaire, CNRS, UMR-6187, University of Poitiers, and

LGCM CHU de Poitiers, Poitiers, France

¹Correspondence: Institut de Physiologie et Biologie Cellulaires, CNRS, UMR-6187, University of Poitiers, 86022 Poitiers, France. E-mail: bruno.constantin{at}univ-poitiers.fr

Calcium mishandling in Duchenne dystrophic muscle suggested that dystrophin, a membrane-associated cytoskeleton protein, might regulate calcium signaling cascade such as calcium influx pathway. It was previously shown that abnormal calcium entries involve uncontrolled stretch-activated currents and store-operated Ca²⁺ currents supported by TRPC1 channels. Moreover, our recent work demonstrated that reintroduction of minidystrophin in dystrophic myotubes restores normal capacitative calcium entries (CCEs). However, until now, no molecular link between the dystrophin complex and calcium entry channels has been described. This study is the first to show by coimmunoprecipitation assays the molecular association of TRPC1 with dystrophin and 1-syntrophin in muscle cells. TRPC1 was also associated with 1-syntrophin in dystrophic muscle cells independently of dystrophin. Furthermore, glutathione S-transferase (GST) pull-down assays showed that TRPC1 binds to the 1-syntrophin PDZ domain. Transfected recombinant 1-syntrophin formed a complex with TRPC1 channels and restored normal CCEs in dystrophic muscle cells. We suggest that normal regulation of CCEs in skeletal muscle depends on the association between TRPC1 channels and 1-syntrophin that may anchor the store-operated channels to the dystrophin-associated protein complex (DAPC). The loss of this molecular association could participate in the calcium alterations observed in dystrophic muscle cells. This study provides a new model for the regulation of calcium influx by interaction with the scaffold of the DAPC in muscle cells. —Vandebrouck, A., Sabourin, J., Rivet, J., Balghi, H., Sebille, S., Kitzis, A., Raymond, G., Cognard, C., Bourmeyster, N., Constantin, B. Regulation of capacitative calcium entries by 1-syntrophin: association of TRPC1 with dystrophin complex and the PDZ domain of 1-syntrophin.

Key Words: dystrophic muscle • store-operated calcium entries • TRP channels • scaffolding proteins

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