Abstract
As the sole Ca2+ entry mechanism in a variety of non-excitable cells, store-operated calcium (SOC) influx is important in Ca2+ signalling and many other cellular processes1,2,3. A calcium-release-activated calcium (CRAC) channel in T lymphocytes is the best-characterized SOC influx channel4,5,6 and is essential to the immune response, sustained activity of CRAC channels being required for gene expression and proliferation7,8,9,10. The molecular identity and the gating mechanism of SOC and CRAC channels have remained elusive. Previously we identified Stim and the mammalian homologue STIM1 as essential components of CRAC channel activation in Drosophila S2 cells and human T lymphocytes11. Here we show that the expression of EF-hand mutants of Stim or STIM1 activates CRAC channels constitutively without changing Ca2+ store content. By immunofluorescence, EM localization and surface biotinylation we show that STIM1 migrates from endoplasmic-reticulum-like sites to the plasma membrane upon depletion of the Ca2+ store. We propose that STIM1 functions as the missing link between Ca2+ store depletion and SOC influx, serving as a Ca2+ sensor that translocates upon store depletion to the plasma membrane to activate CRAC channels.
Similar content being viewed by others
References
Putney, J. W. Jr, Broad, L. M., Braun, F. J., Lievremont, J. P. & Bird, G. S. Mechanisms of capacitative calcium entry. J. Cell Sci. 114, 2223–2229 (2001)
Putney, J. W. Jr Store-operated calcium channels: how do we measure them, and why do we care? [online]. Sci. STKE 2004(243), pe37 (2004) (doi:10.1126/stke.2432004pe37)
Lewis, R. S. Store-operated calcium channels. Adv. Second Messenger Phosphoprotein Res. 33, 279–307 (1999)
Parekh, A. B. & Putney, J. W. Jr Store-operated calcium channels. Physiol. Rev. 85, 757–810 (2005)
Lewis, R. S. & Cahalan, M. D. Mitogen-induced oscillations of cytosolic Ca2+ and transmembrane Ca2+ current in human leukemic T cells. Cell Regul. 1, 99–112 (1989)
Parekh, A. B. & Penner, R. Store depletion and calcium influx. Physiol. Rev. 77, 901–930 (1997)
Lewis, R. S. Calcium signalling mechanisms in T lymphocytes. Annu. Rev. Immunol. 19, 497–521 (2001)
Winslow, M. M., Neilson, J. R. & Crabtree, G. R. Calcium signalling in lymphocytes. Curr. Opin. Immunol. 15, 299–307 (2003)
Feske, S., Giltnane, J., Dolmetsch, R., Staudt, L. M. & Rao, A. Gene regulation mediated by calcium signals in T lymphocytes. Nature Immunol. 2, 316–324 (2001)
Partiseti, M. et al. The calcium current activated by T cell receptor and store depletion in human lymphocytes is absent in a primary immunodeficiency. J. Biol. Chem. 269, 32327–32335 (1994)
Roos, J. et al. STIM1, an essential and conserved component of store-operated Ca2+ channel function. J. Cell Biol. 169, 435–445 (2005)
Yeromin, A. V., Roos, J., Stauderman, K. A. & Cahalan, M. D. A store-operated calcium channel in Drosophila S2 cells. J. Gen. Physiol. 123, 167–182 (2004)
Williams, R. T. et al. Identification and characterization of the STIM (stromal interaction molecule) gene family: coding for a novel class of transmembrane proteins. Biochem. J. 357, 673–685 (2001)
Beckingham, K. Use of site-directed mutations in the individual Ca2+-binding sites of calmodulin to examine Ca2+-induced conformational changes. J. Biol. Chem. 266, 6027–6030 (1991)
Nakayama, S. & Kretsinger, R. H. Evolution of the EF-hand family of proteins. Annu. Rev. Biophys. Biomol. Struct. 23, 473–507 (1994)
Ross, P. E. & Cahalan, M. D. Ca2+ influx pathways mediated by swelling or stores depletion in mouse thymocytes. J. Gen. Physiol. 106, 415–444 (1995)
Prakriya, M. & Lewis, R. S. Potentiation and inhibition of Ca2+ release-activated Ca2+ channels by 2-aminoethyldiphenyl borate (2-APB) occurs independently of IP3 receptors. J. Physiol. (Lond.) 536, 3–19 (2001)
Chung, S. C., McDonald, T. V. & Gardner, P. Inhibition by SK&F 96365 of Ca2+ current, IL-2 production and activation in T lymphocytes. Br. J. Pharmacol. 113, 861–868 (1994)
Eray, M., Matto, M., Kaartinen, M., Andersson, L. & Pelkonen, J. Flow cytometric analysis of apoptotic subpopulations with a combination of annexin V-FITC, propidium iodide, and SYTO 17. Cytometry 43, 134–142 (2001)
Graber, M. N., Alfonso, A. & Gill, D. L. Ca2+ pools and cell growth: arachidonic acid induces recovery of cells growth-arrested by Ca2+ pool depletion. J. Biol. Chem. 271, 883–888 (1996)
Ferrari, D. et al. Endoplasmic reticulum, Bcl-2 and Ca2+ handling in apoptosis. Cell Calcium 32, 413–420 (2002)
Manji, S. S. et al. STIM1: a novel phosphoprotein located at the cell surface. Biochim. Biophys. Acta 1481, 147–155 (2000)
Dolmetsch, R. E. & Lewis, R. S. Signaling between intracellular Ca2+ stores and depletion-activated Ca2+ channels generates [Ca2+]i oscillations in T lymphocytes. J. Gen. Physiol. 103, 365–388 (1994)
Liou, J. et al. STIM is a Ca2+ sensor essential for Ca2+-store-depletion-triggered Ca2+ influx. Curr. Biol. 15, 1235–1241 (2005)
Fanger, C. M. et al. Calcium-activated potassium channels sustain calcium signalling in T lymphocytes, Selective blockers and manipulated channel expression levels. J. Biol. Chem. 276, 12249–12256 (2001)
Acknowledgements
We thank L. Forrest for assistance with cell culture; A. Yeromin, O. Safrina and S. Wei for help with [Ca2+]i imaging; G. Chandy for the use of molecular reagents and laboratory facilities; C. Hughes for providing access to the Amaxa Nucleofector; A. Kolski-Andreaco for the gift of pAc5.1/EGFP; K. Knowlton and D. Summers-Torres from the Department of Medicine, University of California, San Diego for help with deconvolution immunofluorescence microscopy; and P. J. DiGregorio, G. Velicelebi, M. Lioudyno, J. Hall and Y. Li for discussion. Confocal microscopy was performed at the Optical Biology Shared Resource, supported by a Developmental Biology Center and Cancer Center Support Grant at the University of California, Irvine. This work was supported by grants from the National Institutes of Health (to M.D.C. and M.H.E.) and by a fellowship from the Pulmonary Hypertension Association (to Y.Y.).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Supplementary information
Supplementary Notes
Supplementary Methods and Supplementary Figure Legends. (DOC 47 kb)
Supplementary Figures
Supplementary Figures S1–S9. (PDF 2151 kb)
Rights and permissions
About this article
Cite this article
Zhang, S., Yu, Y., Roos, J. et al. STIM1 is a Ca2+ sensor that activates CRAC channels and migrates from the Ca2+ store to the plasma membrane. Nature 437, 902–905 (2005). https://doi.org/10.1038/nature04147
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature04147
- Springer Nature Limited
This article is cited by
-
Role of cytosolic and endoplasmic reticulum Ca2+ in pancreatic beta-cells: pros and cons
Pflügers Archiv - European Journal of Physiology (2024)
-
Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1
Nature Communications (2023)
-
STIM2 regulates NMDA receptor endocytosis that is induced by short-term NMDA receptor overactivation in cortical neurons
Cellular and Molecular Life Sciences (2023)
-
Deletion of TRPC6, an Autism Risk Gene, Induces Hyperexcitability in Cortical Neurons Derived from Human Pluripotent Stem Cells
Molecular Neurobiology (2023)
-
STIM1-Orai1 interaction mediated calcium influx activation contributes to cardiac contractility of insulin-resistant rats
BMC Cardiovascular Disorders (2022)