{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T01:48:00Z","timestamp":1778636880438,"version":"3.51.4"},"reference-count":28,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2021,9,22]],"date-time":"2021-09-22T00:00:00Z","timestamp":1632268800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["142408"],"award-info":[{"award-number":["142408"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"name":"The Arthritis Society","award":["Investigator Award to MJF"],"award-info":[{"award-number":["Investigator Award to MJF"]}]},{"name":"CHU de Qu\u00e9bec Foundation","award":["Funding to MJF"],"award-info":[{"award-number":["Funding to MJF"]}]},{"name":"CHU de Qu\u00e9bec Foundation","award":["Scholarship to JV"],"award-info":[{"award-number":["Scholarship to JV"]}]},{"name":"Fonds Pierre-Borgeat sur l\u2019Arthrites et les Maladies Rhumatismales","award":["Scholarship to JV"],"award-info":[{"award-number":["Scholarship to JV"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>CLEC12A is a myeloid inhibitory receptor that negatively regulates inflammation in mouse models of autoimmune and autoinflammatory arthritis. Reduced CLEC12A expression enhances myeloid cell activation and inflammation in CLEC12A knock-out mice with collagen antibody-induced or gout-like arthritis. Similarly to other C-type lectin receptors, CLEC12A harbours a stalk domain between its ligand binding and transmembrane domains. While it is presumed that the cysteines in the stalk domain have multimerisation properties, their role in CLEC12A expression and\/or signaling remain unknown. We thus used site-directed mutagenesis to determine whether the stalk domain cysteines play a role in CLEC12A expression, internalisation, oligomerisation, and\/or signaling. Mutation of C118 blocks CLEC12A transport through the secretory pathway diminishing its cell-surface expression. In contrast, mutating C130 does not affect CLEC12A cell-surface expression but increases its oligomerisation, inducing ligand-independent phosphorylation of the receptor. Moreover, we provide evidence that CLEC12A dimerisation is regulated in a redox-dependent manner. We also show that antibody-induced CLEC12A cross-linking induces flotillin oligomerisation in insoluble membrane domains in which CLEC12A signals. Taken together, these data indicate that the stalk cysteines in CLEC12A differentially modulate this inhibitory receptor\u2019s expression, oligomerisation and signaling, suggestive of the regulation of CLEC12A in a redox-dependent manner during inflammation.<\/jats:p>","DOI":"10.3390\/ijms221910207","type":"journal-article","created":{"date-parts":[[2021,9,22]],"date-time":"2021-09-22T22:50:48Z","timestamp":1632351048000},"page":"10207","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Regulation of the Expression, Oligomerisation and Signaling of the Inhibitory Receptor CLEC12A by Cysteine Residues in the Stalk Region"],"prefix":"10.3390","volume":"22","author":[{"given":"Julien","family":"Vitry","sequence":"first","affiliation":[{"name":"CHU de Qu\u00e9bec Research Center, Division of Infectious Diseases and Immunology, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"},{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guillaume","family":"Par\u00e9","sequence":"additional","affiliation":[{"name":"CHU de Qu\u00e9bec Research Center, Division of Infectious Diseases and Immunology, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"},{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andr\u00e9a","family":"Murru","sequence":"additional","affiliation":[{"name":"CHU de Qu\u00e9bec Research Center, Division of Infectious Diseases and Immunology, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"},{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xavier","family":"Charest-Morin","sequence":"additional","affiliation":[{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Halim","family":"Maaroufi","sequence":"additional","affiliation":[{"name":"Institute of Integrative Biology and Systems, Laval University, Qu\u00e9bec, QC G1V 0A6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7816-3286","authenticated-orcid":false,"given":"Kenneth R.","family":"McLeish","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Louisville School of Medicine, Louisville, KY 40206, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul H.","family":"Naccache","sequence":"additional","affiliation":[{"name":"CHU de Qu\u00e9bec Research Center, Division of Infectious Diseases and Immunology, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"},{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria J.","family":"Fernandes","sequence":"additional","affiliation":[{"name":"CHU de Qu\u00e9bec Research Center, Division of Infectious Diseases and Immunology, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"},{"name":"Department of Microbiology-Infectious Diseases and Immunology, Faculty of Medicine, Laval University, Qu\u00e9bec, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2159","DOI":"10.1002\/eji.200535628","article-title":"Human MICL (CLEC12A) is differentially glycosylated and is down-regulated following cellular activation","volume":"36","author":"Marshall","year":"2006","journal-title":"Eur. J. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1182\/blood-2005-08-3264","article-title":"Dendritic-cell-associated C-type lectin 2 (DCAL-2) alters dendritic-cell maturation and cytokine production","volume":"107","author":"Chen","year":"2006","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"8443","DOI":"10.1158\/0008-5472.CAN-04-1659","article-title":"C-type lectin-like molecule-1: A novel myeloid cell surface marker associated with acute myeloid leukemia","volume":"64","author":"Bakker","year":"2004","journal-title":"Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2858","DOI":"10.1182\/blood-2004-03-0878","article-title":"KLRL1, a novel killer cell lectinlike receptor, inhibits natural killer cell cytotoxicity","volume":"104","author":"Han","year":"2004","journal-title":"Blood"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"14792","DOI":"10.1074\/jbc.M313127200","article-title":"Identification and Characterization of a Novel Human Myeloid Inhibitory C-type Lectin-like Receptor (MICL) That Is Predominantly Expressed on Granulocytes and Monocytes","volume":"279","author":"Marshall","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1386","DOI":"10.1136\/annrheumdis-2014-206644","article-title":"MICL controls inflammation in rheumatoid arthritis","volume":"75","author":"Redelinghuys","year":"2015","journal-title":"Ann. Rheum. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.immuni.2013.12.015","article-title":"Clec12a Is an Inhibitory Receptor for Uric Acid Crystals that Regulates Inflammation in Response to Cell Death","volume":"40","author":"Neumann","year":"2014","journal-title":"Immunity"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-021-90631-7","article-title":"Expression of the myeloid inhibitory receptor CLEC12A correlates with disease activity and cytokines in early rheumatoid arthritis","volume":"11","author":"Vaillancourt","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"18544","DOI":"10.1073\/pnas.1821351116","article-title":"The uric acid crystal receptor Clec12A potentiates type I interferon responses","volume":"116","author":"Li","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1042\/bss0690059","article-title":"Genomic analysis of C-type lectins","volume":"69","author":"Drickamer","year":"2002","journal-title":"Biochem. Soc. Symp."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1111\/j.1600-065X.1998.tb01185.x","article-title":"The C-type lectin superfamily in the immune system","volume":"163","author":"Weis","year":"1998","journal-title":"Immunol. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1016\/j.molimm.2004.06.013","article-title":"Divergent roles for C-type lectins expressed by cells of the innate immune system","volume":"41","author":"McGreal","year":"2004","journal-title":"Mol. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1080\/07853890600608985","article-title":"C-type lectin-like receptors on myeloid cells","volume":"38","author":"Pyz","year":"2006","journal-title":"Ann. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1016\/S0959-440X(99)00009-3","article-title":"C-type lectin-like domains","volume":"9","author":"Drickamer","year":"1999","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/j.1600-065X.2008.00666.x","article-title":"Immunoreceptor tyrosine-based inhibition motifs: A quest in the past and future","volume":"224","author":"Jaeger","year":"2008","journal-title":"Immunol. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2073","DOI":"10.3390\/biom5032073","article-title":"Splicing Regulation of Pro-Inflammatory Cytokines and Chemokines: At the Interface of the Neuroendocrine and Immune Systems","volume":"5","author":"Shakola","year":"2015","journal-title":"Biomolecules"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Par\u00e9, G., Vitry, J., Merchant, M.L., Vaillancourt, M., Murru, A., Shen, Y., Elowe, S., Lahoud, M.H., Naccache, P.H., and McLeish, K.R. (2021). The Inhibitory Receptor CLEC12A Regulates PI3K-Akt Signaling to Inhibit Neutrophil Activation and Cytokine Release. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.650808"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"R73","DOI":"10.1186\/ar4250","article-title":"Modulation of monosodium urate crystal-induced responses in neutrophils by the myeloid inhibitory C-type lectin-like receptor: Potential therapeutic implications","volume":"15","author":"Marois","year":"2013","journal-title":"Arthritis Res. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"167","DOI":"10.3389\/fnmol.2017.00167","article-title":"Cysteines as Redox Molecular Switches and Targets of Disease","volume":"10","author":"Fra","year":"2017","journal-title":"Front. Mol. Neurosci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.pharmthera.2009.10.007","article-title":"Inhibitory receptors on myeloid cells: New targets for therapy?","volume":"125","author":"Munitz","year":"2010","journal-title":"Pharmacol. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Wiedemann, C., Kumar, A., Lang, A., and Ohlenschl\u00e4ger, O. (2020). Cysteines and Disulfide Bonds as Structure-Forming Units: Insights From Different Domains of Life and the Potential for Characterization by NMR. Front. Chem., 8.","DOI":"10.3389\/fchem.2020.00280"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1182\/blood.V93.5.1464","article-title":"NADPH oxidase: An update","volume":"93","author":"Babior","year":"1999","journal-title":"Blood"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3933","DOI":"10.1242\/jcs.092015","article-title":"The roles of flotillin microdomains\u2013endocytosis and beyond","volume":"124","author":"Otto","year":"2011","journal-title":"J. Cell Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1357","DOI":"10.1093\/glycob\/9.12.1357","article-title":"C-Type lectin-like domains in Caenorhabditis elegans: Predictions from the complete genome sequence","volume":"9","author":"Drickamer","year":"1999","journal-title":"Glycobiology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1111\/j.1365-3083.2011.02665.x","article-title":"The Human C-Type Lectin-Like Receptor CLEC-1 is Upregulated by TGF-\u03b2 and Primarily Localized in the Endoplasmic Membrane Compartment","volume":"75","author":"Sattler","year":"2011","journal-title":"Scand. J. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1089\/104454904322759920","article-title":"Human Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1 Functions as a Dimer in Living Cells","volume":"23","author":"Xie","year":"2004","journal-title":"DNA Cell Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"10860","DOI":"10.1021\/bi201264y","article-title":"Differential Roles of Cysteine Residues in the Cellular Trafficking, Dimerization, and Function of the High-Density Lipoprotein Receptor, SR-BI","volume":"50","author":"Hu","year":"2011","journal-title":"Biochemistry"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1042\/bj20031100","article-title":"Membrane and raft association of reggie-1\/flotillin-2: Role of myristoylation, palmitoylation and oligomerization and induction of filopodia by overexpression","volume":"378","author":"Falkenbach","year":"2004","journal-title":"Biochem. J."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/19\/10207\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:03:24Z","timestamp":1760166204000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/19\/10207"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,22]]},"references-count":28,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["ijms221910207"],"URL":"https:\/\/doi.org\/10.3390\/ijms221910207","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,22]]}}}