Abstract
Mutations in the X-linked gene MAGT1 cause a Congenital Disorder of Glycosylation (CDG), with two distinct clinical phenotypes: a primary immunodeficiency (XMEN disorder) versus intellectual and developmental disability. It was previously established that MAGT1 deficiency abolishes steady-state expression of the immune response protein NKG2D (encoded by KLRK1) in lymphocytes. Here, we show that the reduced steady-state levels of NKG2D are caused by hypoglycosylation of the protein and we pinpoint the exact site that is underglycosylated in MAGT1-deficient patients. Furthermore, we challenge the possibility that supplementation with magnesium restores NKG2D levels and show that the addition of this ion does not significantly improve NKG2D steady-state expression nor does it rescue the hypoglycosylation defect in CRISPR-engineered human cell lines. Moreover, magnesium supplementation of an XMEN patient did not result in restoration of NKG2D expression on the cell surface of lymphocytes. In summary, we demonstrate that in MAGT1-deficient patients, the lack of NKG2D is caused by hypoglycosylation, further elucidating the pathophysiology of XMEN/MAGT1-CDG.
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Abbreviations
- CDG:
-
Congenital disorders of glycosylation
- EH:
-
Endoglycosidase H
- EBV:
-
Epstein-Barr virus
- KO:
-
Knock out
- MFI:
-
Median fluorescence index
- Mg2 + :
-
Magnesium
- OST:
-
Oligosaccharyltransferase
- WT:
-
Wild type
- XMEN:
-
X-linked immunodeficiency with Mg2+ defect: Epstein–Barr virus infection and neoplasia
References
Aebi M (2013) N-linked protein glycosylation in the ER. Biochim Biophys Acta Mol Cell Res 1833:2430–2437. https://doi.org/10.1016/j.bbamcr.2013.04.001
Blommaert E, Péanne R, Cherepanova NA et al (2019) Mutations in MAGT1 lead to a glycosylation disorder with a variable phenotype. Proc Natl Acad Sci 116:9865–9870. https://doi.org/10.1073/pnas.1817815116
Chaigne-Delalande B, Li F-Y, O’Connor GM et al (2013) Mg2+ regulates cytotoxic functions of NK and CD8 T cells in chronic EBV infection through NKG2D. Science 341:186–191
Chauvin SD, Price S, Zou J et al (2021) A double-blind, placebo-controlled, crossover study of mgnesium supplementation in patients with XMEN disease. J Clin Immunol. https://doi.org/10.1007/s10875-021-01137-w
Cherepanova NA, Gilmore R (2016) Mammalian cells lacking either the cotranslational or posttranslocational oligosaccharyltransferase complex display substrate-dependent defects in asparagine linked glycosylation. Sci Rep 6:20946. https://doi.org/10.1038/srep20946
Cherepanova NA, Shrimal S, Gilmore R (2014) Oxidoreductase activity is necessary for N-glycosylation of cysteine-proximal acceptor sites in glycoproteins. J Cell Biol 206:525–539. https://doi.org/10.1083/jcb.201404083
Cherepanova NA, Shrimal S, Gilmore R (2016) N-linked glycosylation and homeostasis of the endoplasmic reticulum. Curr Opin Cell Biol 41:57–65. https://doi.org/10.1016/j.ceb.2016.03.021
Cherepanova NA, Venev SV, Leszyk JD et al (2019) Quantitative glycoproteomics reveals new classes of STT3A- and STT3B-dependent N-glycosylation sites. J Cell Biol 218(8):2782–2796. https://doi.org/10.1083/jcb.201904004
Freeze HH, Chong JX, Bamshad MJ, Ng BG (2014) Solving glycosylation disorders: fundamental approaches reveal complicated pathways. Am J Hum Genet 94:161–175. https://doi.org/10.1016/j.ajhg.2013.10.024
Garrity D, Call ME, Feng J, Wucherpfennig KW (2005) The activating NKG2D receptor assembles in the membrane with two signaling dimers into a hexameric structure. Proc Natl Acad Sci USA 102:7641–7646. https://doi.org/10.1073/pnas.0502439102
Goytain A, Quamme GA (2005) Identification and characterization of a novel mammalian Mg2+ transporter with channel-like properties. BMC Genomics 1:48. https://doi.org/10.1186/1471-2164-6-48
Jaeken J, Péanne R (2017) What is new in CDG? J Inherit Metab Dis 40:569–586. https://doi.org/10.1007/s10545-017-0050-6
Jamieson AM, Diefenbach A, Mcmahon CW et al (2002) The role of the NKG2D immunoreceptor in immune cell activation and natural killing. Immunity 17:19–29
Kelleher DJ, Karaoglu D, Mandon EC, Gilmore R (2003) Oligosaccharyltransferase isoforms that contain different catalytic STT3 subunits have distinct enzymatic properties. Mol Cell 12:101–111. https://doi.org/10.1016/S1097-2765(03)00243-0
Li F-Y, Chaigne-Delalande B, Kanellopoulou C et al (2011) Second messenger role for Mg2+ revealed by human T-cell immunodeficiency. Nature 475:471–476. https://doi.org/10.1038/nature10246
Matsuda-Lennikov M, Biancalana M, Zou J et al (2019) Magnesium transporter 1 (MAGT1) deficiency causes selective defects in N-linked glycosylation and expression of immune-response genes. J Biol Chem 294:13638–13656. https://doi.org/10.1074/jbc.RA119.008903
Münz C (2017) Epstein-Barr virus-specific immune control by innate lymphocytes. Front Immunol 8:1658. https://doi.org/10.3389/fimmu.2017.01658
Pascoal C, Francisco R, Ferro T et al (2020) CDG and immune response: from bedside to bench and back. J Inherit Metab Dis 43:90–124. https://doi.org/10.1002/jimd.12126
Petrescu A-J, Wormald MR, Dwek Ra (2006) Structural aspects of glycomes with a focus on N-glycosylation and glycoprotein folding. Curr Opin Struct Biol 16:600–607. https://doi.org/10.1016/j.sbi.2006.08.007
Raulet DH, Gasser S, Gowen BG et al (2013) Regulation of ligands for the NKG2D activating receptor. Annu Rev Immunol 34:413–441. https://doi.org/10.1146/annurev-immunol-032712-095951
Ravell JC, Chauvin SD, He T, Lenardo M (2020a) An update on XMEN disease. J Clin Immunol 40:671–681. https://doi.org/10.1007/s10875-020-00790-x
Ravell JC, Matsuda-Lennikov M, Chauvin SD et al (2020b) Defective glycosylation and multisystem abnormalities characterize the primary immunodeficiency XMEN disease. J Clin Invest 130:507–522. https://doi.org/10.1172/JCI131116
Wu J, Song Y, Bakker ABH et al (1999) An activating immunoreceptor complex formed by NKG2D and DAP10. Science 285:730–732. https://doi.org/10.1126/science.285.5428.730
Zhou H, Clapham DE (2009) Mammalian MagT1 and TUSC3 are required for cellular magnesium uptake and vertebrate embryonic development. Proc Natl Acad Sci USA 106:15750–15755. https://doi.org/10.1073/pnas.0908332106
Funding
This research was supported by Research Foundation Flanders (FWO): under the frame of E-Rare-3, the ERA-Net for Research on Rare Diseases (ERA-NET Cofund action N°64578) (to GM), a research stay grant (FWO V417818N) (to EB), a senior clinical investigator fellowship (to RS), and GLYCO4DIAG, an International Associated Laboratory grant from National Centre for Scientific Research (CNRS) and FWO (to FF and GM). The work was also supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number GM43768 (to RG), C1 KU Leuven fund (to RS) and by the JAEKEN-THEUNISSEN CDG FUND.
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Designed the experiments EB, NAC, FS, RG, and RS. Performed the experiments EB, NAC, FS, and MW. Analysed the data EB and NAC. EB wrote the manuscript with support from JJ, FF, and GM. Clinical evaluation of patient RS. All authors discussed and revised the manuscript.
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Research on patient cells was approved by the Ethical Committee of the University Hospital of Leuven (approval number S58466).
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Blommaert, E., Cherepanova, N.A., Staels, F. et al. Lack of NKG2D in MAGT1-deficient patients is caused by hypoglycosylation. Hum Genet 141, 1279–1286 (2022). https://doi.org/10.1007/s00439-021-02400-1
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DOI: https://doi.org/10.1007/s00439-021-02400-1