{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T18:36:45Z","timestamp":1793126205437,"version":"4.5.0"},"posted":{"date-parts":[[2026,7,13]]},"group-title":"Cell Biology","reference-count":92,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["R35HL150778"],"award-info":[{"award-number":["R35HL150778"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R35HL183468"],"award-info":[{"award-number":["R35HL183468"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R35HL161177"],"award-info":[{"award-number":["R35HL161177"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["AI175276"],"award-info":[{"award-number":["AI175276"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["AI180128"],"award-info":[{"award-number":["AI180128"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,10,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The Ca\u00b2\u207a release-activated Ca\u00b2\u207a (CRAC) channel mediates store-operated calcium entry (SOCE), a ubiquitous pathway essential for many cell types, including immune cells. Three Orai (Orai1, Orai2, Orai3) proteins constitute the plasma membrane pore-forming units of CRAC channels that are activated by the endoplasmic reticulum (ER) Ca\n                  <jats:sup>2+<\/jats:sup>\n                  -sensing STIM1 and STIM2 proteins when ER Ca\n                  <jats:sup>2+<\/jats:sup>\n                  stores are depleted. Orai1\/2\/3 are differentially expressed across primary cells with discernible differences in their structures and biophysical properties. Further, Orai1 has two alternatively translated isoforms: the long mammalian-specific Orai1\u03b1 and the 63-residue shorter Orai1\u03b2, which is evolutionarily older and conserved across vertebrates. Whether Orai1\u03b1\/1\u03b2\/2\/3 at native expression levels produce unique cytosolic Ca\u00b2\u207a signatures that bias transcriptional responses through effectors like NFAT is unclear. Here we used HEK293 cells engineered to express one native Orai isoform and show that all Orai isoforms couple to NFAT1 and NFAT4 induction. The magnitude of NFAT1\/4 induction tracked the magnitude of SOCE, with the following profile: Orai1\u03b2&gt;Orai1\u03b1&gt;&gt;Orai2&gt;Orai3. Near-native ectopic expression of either Orai1\u03b1 or Orai1\u03b2 in primary murine\n                  <jats:italic>\n                    Orai1\n                    <jats:sup>\u2212\/\u2212<\/jats:sup>\n                  <\/jats:italic>\n                  CD4\u207a T cells restored SOCE, NFAT activation, cytokine production and promoted near identical transcriptional responses enriched for immune activation pathways. An analysis of genetic and clinical data of human individuals showed that homozygous null mutations selectively abolishing Orai1\u03b1 are not associated with disease resembling CRAC channelopathy. Primary T cells from individuals homozygous or heterozygous for an Orai1\u03b1 null mutation showed enhanced, rather than impaired, SOCE and NFAT induction. Collectively, our data indicate that Orai isoforms regulate NFAT activation and downstream transcription primarily through the magnitude of SOCE they support, with Orai1\u03b1 and Orai1\u03b2 being the most potent isoforms coupling SOCE to NFAT responses.\n                <\/jats:p>","DOI":"10.64898\/2026.07.10.737753","type":"posted-content","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T13:05:39Z","timestamp":1783947939000},"source":"Crossref","is-referenced-by-count":0,"title":["Orai1\u03b1 and Orai1\u03b2 more potently promote NFAT activation and transcription than Orai2 and Orai3"],"prefix":"10.64898","author":[{"given":"Ahmed Emam","family":"Abdelnaby","sequence":"first","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yin-Hu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Cory","family":"Benson","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isagani","family":"Perez","sequence":"additional","affiliation":[{"name":"Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boheng","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maxwell","family":"McDermott","sequence":"additional","affiliation":[{"name":"Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miki","family":"Jishage","sequence":"additional","affiliation":[{"name":"Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amal T.","family":"Elhaw","sequence":"additional","affiliation":[{"name":"UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA"},{"name":"Department of Medicine, Division of Malignant Hematology and Medical Oncology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Cruz-Rangel","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raphael","family":"Courjaret","sequence":"additional","affiliation":[{"name":"Calcium Signaling Group, Research Department, Weill Cornell Medicine Qatar, Education City, Qatar Foundation, Doha 24144, Qatar; and Department of Systems and Computational Biomedicine, Weill Cornell Medicine, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdelaziz","family":"Belkadi","sequence":"additional","affiliation":[{"name":"Calcium Signaling Group, Research Department, Weill Cornell Medicine Qatar, Education City, Qatar Foundation, Doha 24144, Qatar; and Department of Systems and Computational Biomedicine, Weill Cornell Medicine, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Yu","sequence":"additional","affiliation":[{"name":"Calcium Signaling Group, Research Department, Weill Cornell Medicine Qatar, Education City, Qatar Foundation, Doha 24144, Qatar; and Department of Systems and Computational Biomedicine, Weill Cornell Medicine, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas S.","family":"Prado","sequence":"additional","affiliation":[{"name":"Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Yuan","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Xin","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam C.","family":"Straub","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco J.","family":"Schopfer","sequence":"additional","affiliation":[{"name":"University of Pittsburgh, Department of Pharmacology and Chemical Biology, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5574-8783","authenticated-orcid":false,"given":"Nadine","family":"Hempel","sequence":"additional","affiliation":[{"name":"UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA"},{"name":"Department of Medicine, Division of Malignant Hematology and Medical Oncology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8371-3345","authenticated-orcid":false,"given":"William F.","family":"Hawse","sequence":"additional","affiliation":[{"name":"Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA"},{"name":"Center for Systems Immunology, University of Pittsburgh, Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David B.","family":"Beck","sequence":"additional","affiliation":[{"name":"Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6215-2411","authenticated-orcid":false,"given":"Khaled","family":"Machaca","sequence":"additional","affiliation":[{"name":"Calcium Signaling Group, Research Department, Weill Cornell Medicine Qatar, Education City, Qatar Foundation, Doha 24144, Qatar; 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