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Published online 9 August 2004. doi:10.1083/jcb.200404079
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 4, 537-548
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Article

 Junctate is a key element in calcium entry induced by activation of InsP3 receptors and/or calcium store depletion

Susan Treves1,2, Clara Franzini-Armstrong4, Luca Moccagatta5, Christophe Arnoult10, Cristiano Grasso5, Adam Schrum2,3, Sylvie Ducreux1, Michael X. Zhu6,7, Katsuhiko Mikoshiba8,9, Thierry Girard1, Sophia Smida-Rezgui10, Michel Ronjat10, and Francesco Zorzato1,5

1 Department of Anesthesiology, University of Basel Kantosspital, Basel, 4031 Switzerland
2 Department of Research, University of Basel Kantosspital, Basel, 4031 Switzerland
3 Department of Transplant Immunology, University of Basel Kantosspital, Basel, 4031 Switzerland
4 Department of Cell and Developmental Biology, University of Pennsylvania, Philadelphia, PA 19104
5 Dipartimento di Medicina Sperimentale e Diagnostica, Università di Ferrara, 44100 Ferrara, Italy
6 Centre for Molecular Neurobiology, Ohio State University, Columbus, OH 43210
7 Department of Neuroscience, Ohio State University, Columbus, OH 43210
8 RIKEN BSI Saitama, ICORP, JST, Tokyo
9 Institute of Medical Science, University of Tokyo, Tokyo, 108-8639 Japan
10 Laboratoire Canaux Calciques, Fonctions et Pathologies, INSERM U607, CEA-Grenoble, France

Address correspondence to Francesco Zorzato, Dipartimento di Medicina Sperimentale e Diagnostica, sez. Patologia Generale Università di Ferrara, Via Borsari 46, 44100 Ferrara, Italy. Tel.: 39-0532-291356 Fax: 39-0532-247278. email: zor{at}unife.it; or fzorzato{at}uhbs.ch

In many cell types agonist-receptor activation leads to a rapid and transient release of Ca2+ from intracellular stores via activation of inositol 1,4,5 trisphosphate (InsP3) receptors (InsP3Rs). Stimulated cells activate store- or receptor-operated calcium channels localized in the plasma membrane, allowing entry of extracellular calcium into the cytoplasm, and thus replenishment of intracellular calcium stores. Calcium entry must be finely regulated in order to prevent an excessive intracellular calcium increase. Junctate, an integral calcium binding protein of endo(sarco)plasmic reticulum membrane, (a) induces and/or stabilizes peripheral couplings between the ER and the plasma membrane, and (b) forms a supramolecular complex with the InsP3R and the canonical transient receptor potential protein (TRPC) 3 calcium entry channel. The full-length protein modulates both agonist-induced and store depletion–induced calcium entry, whereas its NH2 terminus affects receptor-activated calcium entry. RNA interference to deplete cells of endogenous junctate, knocked down both agonist-activated calcium release from intracellular stores and calcium entry via TRPC3. These results demonstrate that junctate is a new protein involved in calcium homeostasis in eukaryotic cells.

Key Words: calcium homeostasis; IP3R; calcium entry channel; calcium binding protein; ER

Abbreviations used in this paper: 2-APB, 2-aminoethyl diphenylborate; EYFP, enhanced YFP; InsP3, inositol 1,4,5 trisphosphate; InsP3R, InsP3 receptor; RNAi; RNA interference; SERCA, sarcoplasmic and ER Ca2+ATPase; t-BuBHQ, Di-tert-butylhydroquinone; TRPC, canonical transient receptor potential protein.


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