The channel Orai1 requires Ca2+ store depletion in the endoplasmic reticulum and an interaction with the Ca2+ sensor STIM1 to mediate Ca2+ signaling. Alterations in Orai1-mediated Ca2+ influx have been linked to several pathological conditions including immunodeficiency, tubular myopathy, and cancer. We screened large-scale cancer genomics data sets for dysfunctional Orai1 mutants. Five of the identified Orai1 mutations resulted in constitutively active gating and transcriptional activation. Our analysis showed that certain Orai1 mutations were clustered in the transmembrane 2 helix surrounding the pore, which is a trigger site for Orai1 channel gating. Analysis of the constitutively open Orai1 mutant channels revealed two fundamental gates that enabled Ca2+ influx: Arginine side chains were displaced so they no longer blocked the pore, and a chain of water molecules formed in the hydrophobic pore region. Together, these results enabled us to identify a cluster of Orai1 mutations that trigger Ca2+ permeation associated with gene transcription and provide a gating mechanism for Orai1.

Center for Nanobiology and Structural Biology Institute of Microbiology Academy of Sciences of the Czech Republic Nové Hrady CZ 373 33 Czech Republic

Center of New Technologies University of Warsaw Warsaw 02 097 Poland

Department of Biophysics Center for Integrative Physiology and Molecular Medicine Medical Faculty Saarland University Homburg D 66421 Germany

Faculty of Sciences University of South Bohemia Nové Hrady CZ 373 33 Czech Republic

Institute for Biophysics Medical University of Graz Graz A 8010 Austria

Institute for Experimental and Clinical Cell Therapy Paracelsus Medical University Salzburg A 5020 Austria

Institute for Molecular Biosciences Karl Franzens University Graz Graz A 8010 Austria

Institute of Biophysics JKU Life Science Center Johannes Kepler University Linz Linz A 4020 Austria

Max Delbrück Center for Molecular Medicine in the Helmholtz Association Cardiovascular and Metabolic Sciences Berlin D 13125 Germany

Molecular Physiology Institute of Cardiovascular Physiology University Medical Center Georg August University Göttingen Göttingen Niedersachsen 37073 Germany

University of Chicago Chicago IL 60637 USA

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Sci Signal. 2017 Nov 28;10(507):eaaq0618. doi: 10.1126/scisignal.aaq0618. PubMed

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CRAC channel opening is determined by a series of Orai1 gating checkpoints in the transmembrane and cytosolic regions

Blockage of Store-Operated Ca2+ Influx by Synta66 is Mediated by Direct Inhibition of the Ca2+ Selective Orai1 Pore

Luminal STIM1 Mutants that Cause Tubular Aggregate Myopathy Promote Autophagic Processes