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Store-operated cyclic AMP signalling mediated by STIM1

Nature Cell Biology volume 11, pages 433–442 (2009)Cite this article

Abstract

Depletion of Ca2+ from the endoplasmic reticulum (ER) results in activation of plasma membrane Ca2+ entry channels. This 'store-operated' process requires translocation of a transmembrane ER Ca2+ sensor protein, stromal interaction molecule 1 (STIM1), to sites closely apposed to Ca2+ channels at the cell surface. However, it is not known whether a reduction in Ca2+ stores is coupled to other signalling pathways by this mechanism. We found that lowering the concentration of free Ca2+ in the ER, independently of the cytosolic Ca2+ concentration, also led to recruitment of adenylyl cyclases. This resulted in enhanced cAMP accumulation and PKA activation, measured using FRET-based cAMP indicators. Translocation of STIM1 was required for efficient coupling of ER Ca2+ depletion to adenylyl cyclase activity. We propose the existence of a pathway (store-operated cAMP signalling or SOcAMPS) in which the content of internal Ca2+ stores is directly connected to cAMP signalling through a process that involves STIM1.

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Figure 1: Chelation of Ca2+ within internal stores of NCM460 cells induces STIM1 translocation, cAMP signalling and PKA phosphorylation.
Figure 2: Depletion of internal Ca2+ stores with ionomycin enhances cAMP signalling, PKA phosphorylation and expression of ICER in NCM460 cells.
Figure 3: Effects of store depletion on intracellular cAMP in the presence of extracellular Ca2+ in NCM460 cells.
Figure 4: Store depletion-induced cAMP signalling results from activation of ACs and not inhibition of PDEs.
Figure 5: STIM1 expression is required for store-operated cAMP signalling.
Figure 6: Translocation of STIM protein is necessary to initiate store-operated cAMP signalling.

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Acknowledgements

We are grateful to the following individuals for their kind gifts of plasmids: Kees Jalink for the Epac sensor, Roger Tsien for mCherry, Jin Zhang for AKAR3, Marie Dziadek and Lorna Johnstone for STIM1 and STIM2, Masaru Okabe for pCX–EGFP, and Tobias Meyer for YFP–STIM1, YFP–STIM1D76A, and YFP–STIM2 constructs. We thank Jessica Roy for technical assistance and Drs. Dheeraj Pelluru, Eberhard Frömter and Raj K. Goyal for helpful comments. This study was supported by a Merit Review award from the Department of Veteran's Affairs (A.M.H.) and by an NIH Center grant from the Harvard Digestive Diseases Center (to A.M.H.). K.L. is the recipient of an American Heart Association Postdoctoral Fellowship award.

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  1. Isabella Maiellaro

    Present address: Current address: Dipartimento di Fisiologia Generale ed Ambientale, Universitá degli Studi di Bari, Bari, Italy 70126.,

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  1. VA Boston Healthcare System and the Department of Surgery, Brigham and Women's Hospital and Harvard Medical School, 1400 VFW Parkway, West Roxbury, 02132, Massachusetts, USA

    Konstantinos Lefkimmiatis, Meera Srikanthan, Isabella Maiellaro, Silvana Curci & Aldebaran M. Hofer

  2. INCELL Corporation LLC, San Antonio, 78249, Texas, USA

    Mary Pat Moyer

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Contributions

K.L., I.M. and A.M.H. performed experiments; K.L., A.M.H., and S.C. designed experiments, and along with M.S., analyzed data. M.P.M. provided reagents. A.M.H. wrote the manuscript.

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Correspondence to Aldebaran M. Hofer.

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M.P.M. holds partial ownership of INCELL Corporation, which sells the M3:10 tissue culture medium used to grow NCM460 cells. The other authors declare no competing financial interests.

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Lefkimmiatis, K., Srikanthan, M., Maiellaro, I. et al. Store-operated cyclic AMP signalling mediated by STIM1. Nat Cell Biol 11, 433–442 (2009). https://doi.org/10.1038/ncb1850

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