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The Endoplasmic Reticulum Stress Sensor IRE1α Regulates the UV DNA Repair Response through the Control of Intracellular Calcium Homeostasis

  • Jeongin Son
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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  • Saie Mogre
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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  • Fiona E. Chalmers
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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  • Jack Ibinson
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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  • Stephen Worrell
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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  • Adam B. Glick
    Correspondence
    Correspondence: Adam B. Glick, Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, 101 Life Sciences Building, University Park, Pennsylvania 16802, USA.
    Affiliations
    Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences, The Pennsylvania State University, Pennsylvania, USA
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Published:November 19, 2021DOI:https://doi.org/10.1016/j.jid.2021.11.010
      The unfolded protein response is activated by UVB irradiation, but the role of a key mediator, IRE1α, is not clear. In this study, we show that mice with an epidermal IRE1α deletion are sensitized to UV with increased apoptosis, rapid loss of UV-induced cyclopyrimidine dimer‒positive keratinocytes, and sloughing of the epidermis. In vitro, Ire-deficient keratinocytes have increased UVB sensitivity, reduced cyclopyrimidine dimer repair, and reduced accumulation of γH2AX and phosphorylated ATR, suggesting defective activation of nucleotide excision repair. Knockdown of XBP1 or pharmacologic inhibition of the IRE1α ribonuclease did not phenocopy Ire deficiency. The altered UV response was linked to elevated intracellular calcium levels and ROS, and this was due to dysregulation of the endoplasmic reticulum calcium channel InsP3R. Pharmacologic, genetic, and biochemical studies linked the regulation of the Ins3PR, intracellular calcium, and normal UV DNA damage response to CIB1 and the IRE1α‒TRAF2‒ASK1 complex. These results suggest a model where IRE1α activation state drives CIB1 binding either to the InsP3R or ASK1 to regulate endoplasmic reticulum calcium efflux, ROS, and DNA repair responses after UV irradiation.

      Abbreviations:

      [Ca2+]i (intracellular calcium concentration), CPD (cycloprimidine dimer), DDR (DNA damage response), ER (endoplasmic reticulum), KC (keratinocyte), KD (knockdown), p-ATR (phosphorylated ATR), RIDD (regulated IRE1α-dependent decay), RNase (ribonuclease), UPR (unfolded protein response)
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      References

        • Assefa Z.
        • Garmyn M.
        • Bouillon R.
        • Merlevede W.
        • Vandenheede J.R.
        • Agostinis P.
        Differential stimulation of ERK and JNK activities by ultraviolet B irradiation and epidermal growth factor in human keratinocytes.
        J Invest Dermatol. 1997; 108: 886-891
        • Bankhead P.
        • Loughrey M.B.
        • Fernández J.A.
        • Dombrowski Y.
        • McArt D.G.
        • Dunne P.D.
        • et al.
        QuPath: open source software for digital pathology image analysis.
        Sci Rep. 2017; 7: 16878
        • Blazanin N.
        • Son J.
        • Craig-Lucas A.B.
        • John C.L.
        • Breech K.J.
        • Podolsky M.A.
        • et al.
        ER stress and distinct outputs of the IRE1α RNase control proliferation and senescence in response to oncogenic Ras.
        Proc Natl Acad Sci USA. 2017; 114: 9900-9905
        • Boukamp P.
        • Petrussevska R.T.
        • Breitkreutz D.
        • Hornung J.
        • Markham A.
        • Fusenig N.E.
        Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.
        J Jcell Biol. 1988; 106: 761-771
        • Brown M.
        • Strudwick N.
        • Suwara M.
        • Sutcliffe L.K.
        • Mihai A.D.
        • Ali A.A.
        • et al.
        An initial phase of JNK activation inhibits cell death early in the endoplasmic reticulum stress response.
        J Cell Sci. 2016; 129: 2317-2328
        • Castillo K.
        • Rojas-Rivera D.
        • Lisbona F.
        • Caballero B.
        • Nassif M.
        • Court F.A.
        • et al.
        BAX inhibitor-1 regulates autophagy by controlling the IRE1α branch of the unfolded protein response.
        EMBO J. 2017; 36: 1640
        • Chalmers F.
        • Mogre S.
        • Son J.
        • Blazanin N.
        • Glick A.B.
        The multiple roles of the unfolded protein response regulator IRE1α in cancer.
        Mol Carcinog. 2019; 58: 1623-1630
        • Chen X.
        • Cubillos-Ruiz J.R.
        Endoplasmic reticulum stress signals in the tumour and its microenvironment.
        Nat Rev Cancer. 2021; 21: 71-88
        • Collier A.E.
        • Wek R.C.
        • Spandau D.F.
        Translational repression protects human keratinocytes from UVB-induced apoptosis through a discordant eIF2 kinase stress response.
        J Invest Dermatol. 2015; 135: 2502-2511
        • Corazzari M.
        • Rapino F.
        • Ciccosanti F.
        • Giglio P.
        • Antonioli M.
        • Conti B.
        • et al.
        Oncogenic BRAF induces chronic ER stress condition resulting in increased basal autophagy and apoptotic resistance of cutaneous melanoma.
        Cell Death Differ. 2015; 22: 946-958
        • Dufey E.
        • Bravo-San Pedro J.M.
        • Eggers C.
        • González-Quiroz M.
        • Urra H.
        • Sagredo A.I.
        • et al.
        Genotoxic stress triggers the activation of IRE1α-dependent RNA decay to modulate the DNA damage response.
        Nat Commun. 2020; 11: 2401
        • Farrukh M.R.
        • Nissar U.A.
        • Afnan Q.
        • Rafiq R.A.
        • Sharma L.
        • Amin S.
        • et al.
        Oxidative stress mediated Ca(2+) release manifests endoplasmic reticulum stress leading to unfolded protein response in UV-B irradiated human skin cells.
        J Dermatol Sci. 2014; 75: 24-35
        • González-Quiroz M.
        • Blondel A.
        • Sagredo A.
        • Hetz C.
        • Chevet E.
        • Pedeux R.
        When endoplasmic reticulum proteostasis meets the DNA damage response.
        Trends Cell Biol. 2020; 30: 881-891
        • Gumeni S.
        • Evangelakou Z.
        • Gorgoulis V.G.
        • Trougakos I.P.
        Proteome stability as a key factor of genome integrity.
        Int J Mol Sci. 2017; 18: 2036
        • Hanasoge S.
        • Ljungman M.
        H2AX phosphorylation after UV irradiation is triggered by DNA repair intermediates and is mediated by the ATR kinase.
        Carcinogenesis. 2007; 28: 2298-2304
        • Hogan K.A.
        • Ravindran A.
        • Podolsky M.A.
        • Glick A.B.
        The TGFβ1 pathway is required for NFκB dependent gene expression in mouse keratinocytes.
        Cytokine. 2013; 64: 652-659
        • Hung K.F.
        • Sidorova J.M.
        • Nghiem P.
        • Kawasumi M.
        The 6–4 photoproduct is the trigger of UV-induced replication blockage and ATR activation.
        Proc Natl Acad Sci USA. 2020; 117: 12806-12816
        • Indra A.K.
        • Warot X.
        • Brocard J.
        • Bornert J.M.
        • Xiao J.H.
        • Chambon P.
        • et al.
        Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases.
        Nucleic Acids Res. 1999; 27: 4324-4327
        • Iwawaki T.
        • Akai R.
        • Yamanaka S.
        • Kohno K.
        Function of IRE1 alpha in the placenta is essential for placental development and embryonic viability.
        Proc Natl Acad Sci USA. 2009; 106: 16657-16662
        • Kawachi Y.
        • Xu X.
        • Taguchi S.
        • Sakurai H.
        • Nakamura Y.
        • Ishii Y.
        • et al.
        Attenuation of UVB-induced sunburn reaction and oxidative DNA damage with no alterations in UVB-induced skin carcinogenesis in Nrf2 gene-deficient mice.
        J Invest Dermatol. 2008; 128: 1773-1779
        • Liu C.
        • Yan D.Y.
        • Wang C.
        • Ma Z.
        • Deng Y.
        • Liu W.
        • et al.
        IRE1 signaling pathway mediates protective autophagic response against manganese-induced neuronal apoptosis in vivo and in vitro.
        Sci Total Environ. 2020; 712: 136480
        • Madronich S.
        • de Gruijl F.R.
        Skin cancer and UV radiation.
        Nature. 1993; 366: 23
        • Masaki H.
        • Izutsu Y.
        • Yahagi S.
        • Okano Y.
        Reactive oxygen species in HaCaT keratinocytes after UVB irradiation are triggered by intracellular Ca(2+) levels.
        J Investig Dermatol Symp Proc. 2009; 14: 50-52
        • Mera K.
        • Kawahara K.
        • Tada K.
        • Kawai K.
        • Hashiguchi T.
        • Maruyama I.
        • et al.
        ER signaling is activated to protect human HaCaT keratinocytes from ER stress induced by environmental doses of UVB.
        Biochem Biophys Res Commun. 2010; 397: 350-354
        • Oslowski C.M.
        • Hara T.
        • O'Sullivan-Murphy B.
        • Kanekura K.
        • Lu S.
        • Hara M.
        • et al.
        Thioredoxin-interacting protein mediates ER stress-induced β cell death through initiation of the inflammasome.
        Cell Metab. 2012; 16: 265-273
        • Pinton P.
        • Giorgi C.
        • Siviero R.
        • Zecchini E.
        • Rizzuto R.
        Calcium and apoptosis: ER-mitochondria Ca2+ transfer in the control of apoptosis.
        Oncogene. 2008; 27: 6407-6418
        • Ravindran A.
        • Mohammed J.
        • Gunderson A.J.
        • Cui X.
        • Glick A.B.
        Tumor-promoting role of TGFβ1 signaling in ultraviolet B-induced skin carcinogenesis is associated with cutaneous inflammation and lymph node migration of dermal dendritic cells.
        Carcinogenesis. 2014; 35: 959-966
        • Ray A.
        • Blevins C.
        • Wani G.
        • Wani A.A.
        ATR- and ATM-mediated DNA damage response is dependent on excision repair assembly during G1 but not in S phase of cell cycle.
        PLoS One. 2016; 11e0159344
        • Riaz T.A.
        • Junjappa R.P.
        • Handigund M.
        • Ferdous J.
        • Kim H.R.
        • Chae H.J.
        Role of endoplasmic reticulum stress sensor IRE1α in cellular physiology, calcium, ROS Signaling, and metaflammation.
        Cells. 2020; 9: 1160
        • Roux K.J.
        • Kim D.I.
        • Burke B.
        • May D.G.
        BioID: a screen for protein-protein interactions.
        Curr Protoc Protein Sci. 2018; 91: 19.23.1-19.23.15
        • Seo M.D.
        • Enomoto M.
        • Ishiyama N.
        • Stathopulos P.B.
        • Ikura M.
        Structural insights into endoplasmic reticulum stored calcium regulation by inositol 1,4,5-trisphosphate and ryanodine receptors.
        Biochim Biophys Acta. 2015; 1853: 1980-1991
        • Seo S.H.
        • Kim S.E.
        • Lee S.E.
        ER stress induced by ER calcium depletion and UVB irradiation regulates tight junction barrier integrity in human keratinocytes.
        J Dermatol Sci. 2020; 98: 41-49
        • Shaj K.
        • Hutcherson R.J.
        • Kemp M.G.
        ATR kinase activity limits mutagenesis and promotes the clonogenic survival of quiescent human keratinocytes exposed to UVB radiation.
        Photochem Photobiol. 2020; 96: 105-112
        • Shiizaki S.
        • Naguro I.
        • Ichijo H.
        Activation mechanisms of ASK1 in response to various stresses and its significance in intracellular signaling.
        Adv Biol Regul. 2013; 53: 135-144
        • Son S.M.
        • Byun J.
        • Roh S.E.
        • Kim S.J.
        • Mook-Jung I.
        Reduced IRE1α mediates apoptotic cell death by disrupting calcium homeostasis via the InsP3 receptor.
        Cell Death Dis. 2014; 5: e1188
        • Tan H.Y.
        • Wang N.
        • Tsao S.W.
        • Che C.M.
        • Yuen M.F.
        • Feng Y.
        IRE1α inhibition by natural compound genipin on tumour associated macrophages reduces growth of hepatocellular carcinoma.
        Oncotarget. 2016; 7: 43792-43804
        • Urano F.
        • Wang X.
        • Bertolotti A.
        • Zhang Y.
        • Chung P.
        • Harding H.P.
        • et al.
        Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1.
        Science. 2000; 287: 664-666
        • Wang M.
        • Kaufman R.J.
        The impact of the endoplasmic reticulum protein-folding environment on cancer development.
        Nat Rev Cancer. 2014; 14: 581-597
        • Zeng T.
        • Peng L.
        • Chao H.
        • Xi H.
        • Fu B.
        • Wang Y.
        • et al.
        IRE1α-TRAF2-ASK1 complex-mediated endoplasmic reticulum stress and mitochondrial dysfunction contribute to CXC195-induced apoptosis in human bladder carcinoma T24 cells.
        Biochem Biophys Res Commun. 2015; 460: 530-536

      Supplementary References

        • Bankhead P.
        • Loughrey M.B.
        • Fernández J.A.
        • Dombrowski Y.
        • McArt D.G.
        • Dunne P.D.
        • et al.
        QuPath: open source software for digital pathology image analysis.
        Sci Rep. 2017; 7: 16878
        • Blazanin N.
        • Son J.
        • Craig-Lucas A.B.
        • John C.L.
        • Breech K.J.
        • Podolsky M.A.
        • et al.
        ER stress and distinct outputs of the IRE1α RNase control proliferation and senescence in response to oncogenic Ras.
        Proc Natl Acad Sci USA. 2017; 114: 9900-9905
        • Franken N.A.
        • Rodermond H.M.
        • Stap J.
        • Haveman J.
        • van Bree C.
        Clonogenic assay of cells in vitro.
        Nat Protoc. 2006; 1: 2315-2319
        • Hogan K.A.
        • Ravindran A.
        • Podolsky M.A.
        • Glick A.B.
        The TGFβ1 pathway is required for NFκB dependent gene expression in mouse keratinocytes.
        Cytokine. 2013; 64: 652-659
        • Kobayashi D.
        • Ohkubo S.
        • Nakahata N.
        Contribution of extracellular signal-regulated kinase to UTP-induced interleukin-6 biosynthesis in HaCaT keratinocytes.
        J Pharmacol Sci. 2006; 102: 368-376
        • Ravindran A.
        • Mohammed J.
        • Gunderson A.J.
        • Cui X.
        • Glick A.B.
        Tumor-promoting role of TGFβ1 signaling in ultraviolet B-induced skin carcinogenesis is associated with cutaneous inflammation and lymph node migration of dermal dendritic cells.
        Carcinogenesis. 2014; 35: 959-966
        • Spohn D.
        • Rössler O.G.
        • Philipp S.E.
        • Raubuch M.
        • Kitajima S.
        • Griesemer D.
        • et al.
        Thapsigargin induces expression of activating transcription factor 3 in human keratinocytes involving Ca2+ Ions and c-Jun N-terminal protein kinase.
        Mol Pharmacol. 2010; 78: 865-876
        • Tsutsumi M.
        • Kitahata H.
        • Nakata S.
        • Sanno Y.
        • Nagayama M.
        • Denda M.
        Mathematical analysis of intercellular calcium propagation induced by adenosine triphosphate.
        Skin Res Technol. 2010; 16: 146-150