Abstract
Antigen stimulation of immune cells triggers Ca2+ entry through Ca2+ release-activated Ca2+ (CRAC) channels, promoting the immune response to pathogens by activating the transcription factor NFAT. We have previously shown that cells from patients with one form of hereditary severe combined immune deficiency (SCID) syndrome are defective in store-operated Ca2+ entry and CRAC channel function. Here we identify the genetic defect in these patients, using a combination of two unbiased genome-wide approaches: a modified linkage analysis with single-nucleotide polymorphism arrays, and a Drosophila RNA interference screen designed to identify regulators of store-operated Ca2+ entry and NFAT nuclear import. Both approaches converged on a novel protein that we call Orai1, which contains four putative transmembrane segments. The SCID patients are homozygous for a single missense mutation in ORAI1, and expression of wild-type Orai1 in SCID T cells restores store-operated Ca2+ influx and the CRAC current (ICRAC). We propose that Orai1 is an essential component or regulator of the CRAC channel complex.
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Acknowledgements
We thank S. Ehl, I. Barlan, G. Tuncman and T. Akkoc for facilitating the contact with the SCID patients' families; B. Mathey-Prevot, N. Perrimon, and staff at the Drosophila RNAi Screening Centre at Harvard Medical School for valuable assistance with the screens; M. Komarinski for software support (Genehunter 2.1r5); J. Nardone for help with sequence analysis; S. Sharma for providing the S2R + cell line stably transduced with NFAT–GFP; and D. Neems for technical assistance. This work was funded by grants from the National Institutes of Health to A.R., R.S.L and S.F., and an Immune Deficiency Foundation grant to S.F. Author Contributions S.F. was responsible for all experiments involving genetic and functional analysis of SCID patients, and was assisted in these experiments by S.-H.P. Y.G. designed and implemented the genome-wide Drosophila RNA screen that identified dOrai, with assistance from S.S. M.P. and R.S.L. analysed the electrophysiological properties of calcium currents in Orai1-reconstituted SCID T cells provided by S.F. M.D. advised on the design of the linkage mapping screen and analysis of linkage data, which was carried out by B.T. P.H. and A.R. provided advice and overall direction, and supervised project planning and execution.
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Feske, S., Gwack, Y., Prakriya, M. et al. A mutation in Orai1 causes immune deficiency by abrogating CRAC channel function. Nature 441, 179–185 (2006). https://doi.org/10.1038/nature04702
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DOI: https://doi.org/10.1038/nature04702
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