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Stress granules support cell survival by sequestering components of the translational machinery, suppressing pro-apoptotic signaling, and conserving cellular resources. O-GlcNAcylation, a dynamic modification of cytoplasmic proteins, is an important regulator of stress responses, including stress-granule assembly. O-GlcNAcylation is protective in acute myocardial stress; however, whether O-GlcNAcylation regulates stress granules to protect cardiomyocytes remains unclear. Here, we investigated whether O-GlcNAcylation regulates stress granule formation in cardiomyocytes exposed to sodium arsenite stress or ischemia\/reperfusion (I\/R) injury. G3BP1 is an RNA-binding protein that is essential in stress granule formation. Using an EGFP\u2013G3BP1 reporter, we found that OGT, the enzyme responsible for installing O-GlcNAc on target proteins, is necessary for efficient stress-granule formation in cardiomyocytes. Importantly, we identified T268 of G3BP1 as an O-GlcNAcylation site in these cells and demonstrated by mutagenesis its functional importance for stress granule assembly. In primary cardiomyocytes subjected to simulated I\/R, stress granules formed transiently during ischemia, and this response was decreased by OGT-knockdown. Similarly, stress granules were transiently detected at the ischemic phase of I\/R injury, and their numbers were reduced in OGT-deficient hearts. Furthermore, pretreatment with sodium arsenite before I\/R was cardioprotective. Likewise, pretreatment with sodium arsenite protected cells from staurosporine-induced death, and this effect was decreased by OGT knockdown. Together, these findings identify OGT and O-GlcNAcylation as important regulators of stress-granule assembly in cardiomyocytes, define G3BP1 as an O-GlcNAc-modified effector, and demonstrate that stress granules can act as cytoprotective mediators during acute cardiac injury.<\/jats:p>","DOI":"10.1042\/bcj20250300","type":"journal-article","created":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T14:03:52Z","timestamp":1787234632000},"page":"1947-1977","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Loss of O-GlcNAc transferase activity impairs the dynamic formation of protective stress granules in ischemic cardiomyocytes"],"prefix":"10.1042","volume":"483","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-8170-805X","authenticated-orcid":false,"given":"Hannah\u00a0C.","family":"Swimm","sequence":"first","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenxi","family":"Zhang","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4840-8789","authenticated-orcid":false,"given":"Yashas","family":"Mallikarjun","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-5493-5062","authenticated-orcid":false,"given":"Rohit","family":"Satish","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6897-0042","authenticated-orcid":false,"given":"Andres\u00a0J.","family":"Medina","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-5921-1160","authenticated-orcid":false,"given":"Sabrina","family":"Lo","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hisham","family":"Qureshi","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0333-8975","authenticated-orcid":false,"given":"Aidan\u00a0J.","family":"Dunphy","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0252-4716","authenticated-orcid":false,"given":"Deepthi","family":"Ashok","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Agnes","family":"Sidor","sequence":"additional","affiliation":[{"name":"1Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, 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