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Structural, biochemical, and genetic data demonstrate that NAIP proteins are receptors for bacterial ligands, while NLRC4 is a downstream adaptor that multimerizes with NAIPs to form an inflammasome. NLRC4 has also been proposed to suppress tumor growth, though the underlying mechanism is unknown. Further, NLRC4 is phosphorylated on serine 533, which was suggested to be critical for its function. In the absence of S533 phosphorylation, it was proposed that another inflammasome protein, NLRP3, can induce NLRC4 activation. We generated a new Nlrc4-deficient mouse line and mice with S533D phosphomimetic or S533A nonphosphorylatable NLRC4. Using these models in vivo and in vitro, we fail to observe a requirement for phosphorylation in NLRC4 inflammasome function. Furthermore, we find no role for NLRP3 in NLRC4 function, or for NLRC4 in a model of melanoma. These results clarify our understanding of the mechanism and biological functions of NAIP\/NLRC4 activation.<\/jats:p>","DOI":"10.1084\/jem.20191736","type":"journal-article","created":{"date-parts":[[2020,4,2]],"date-time":"2020-04-02T02:21:27Z","timestamp":1585794087000},"update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":64,"title":["NLRC4 inflammasome activation is NLRP3- and phosphorylation-independent during infection and does not protect from melanoma"],"prefix":"10.1084","volume":"217","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0175-080X","authenticated-orcid":true,"given":"Jeannette L.","family":"Tenthorey","sequence":"first","affiliation":[{"name":"Molecular and Cell Biology Department, Immunology and Pathogenesis Division, and Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto A.","family":"Chavez","sequence":"additional","affiliation":[{"name":"Molecular and Cell Biology Department, Immunology and Pathogenesis Division, and Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thornton W.","family":"Thompson","sequence":"additional","affiliation":[{"name":"Molecular and Cell Biology Department, Immunology and Pathogenesis Division, and Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine A.","family":"Deets","sequence":"additional","affiliation":[{"name":"Molecular and Cell Biology Department, Immunology and Pathogenesis Division, and Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6686-3912","authenticated-orcid":true,"given":"Russell E.","family":"Vance","sequence":"additional","affiliation":[{"name":"Molecular and Cell Biology Department, Immunology and Pathogenesis Division, and Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 1"},{"name":"Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1897-5693","authenticated-orcid":true,"given":"Isabella","family":"Rauch","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR 3"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2020,4,27]]},"reference":[{"key":"2023072611032304300_bib1","doi-asserted-by":"publisher","first-page":"1045","DOI":"10.1084\/jem.20100050","article-title":"The NLRP3 inflammasome functions as a negative regulator of tumorigenesis during colitis-associated cancer","volume":"207","author":"Allen","year":"2010","journal-title":"J. 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