Modification of STIM1 by O-linked N-Acetylglucosamine (O-GlcNAc) Attenuates Store-operated Calcium Entry in Neonatal Cardiomyocytes

Store-operated calcium entry (SOCE) is a major Ca2+ signaling pathway responsible for regulating numerous transcriptional events. In cardiomyocytes SOCE has been shown to play an important role in regulating hypertrophic signaling pathways, including nuclear translocation of NFAT. Acute activation of pathways leading to O-GlcNAc synthesis have been shown to impair SOCE-mediated transcription and in diabetes, where O-GlcNAc levels are chronically elevated, cardiac hypertrophic signaling is also impaired. Therefore the goal of this study was to determine whether changes in cardiomyocyte O-GlcNAc levels impaired the function of STIM1, a widely recognized mediator of SOCE. We demonstrated that acute activation of SOCE in neonatal cardiomyocytes resulted in STIM1 puncta formation, which was inhibited in a dose-dependent manner by increasing O-GlcNAc synthesis with glucosamine or inhibiting O-GlcNAcase with thiamet-G. Glucosamine and thiamet-G also inhibited SOCE and were associated with increased O-GlcNAc modification of STIM1. These results suggest that activation of cardiomyocyte O-GlcNAcylation attenuates SOCE via STIM1 O-GlcNAcylation and that this may represent a new mechanism by which increased O-GlcNAc levels regulate Ca2+-mediated events in cardiomyocytes. Further, since SOCE is a fundamental mechanism underlying Ca2+ signaling in most cells and tissues, it is possible that STIM1 represents a nexus linking protein O-GlcNAcylation with Ca2+-mediated transcription. Background: Increased cellular O-GlcNAc levels decrease store-operated Ca2+ entry (SOCE), however, the mechanism is not understood. STIM1 regulates SOCE, but effect of O-GlcNAc on STIM1 function is not known. Results: Increased cardiomyocyte O-GlcNAcylation attenuated STIM1 puncta formation, SOCE and increased O-GlcNAc modification of STIM1. Conclusion:O-GlcNAc modification of STIM1 plays a key role in regulating SOCE. Significance: Protein O-GlcNAcylation regulates SOCE, a central Ca2+ signaling pathway.