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Mol. Biosci."],"abstract":"<jats:p>The modification of proteins by O-linked \u03b2-<jats:italic>N<\/jats:italic>-acetylglucosamine (O-GlcNAc) is associated with the regulation of numerous cellular processes. Despite the importance of O-GlcNAc in mediating cellular function our understanding of the mechanisms that regulate O-GlcNAc levels is limited. One factor known to regulate protein O-GlcNAc levels is nutrient availability; however, the fact that nutrient deficient states such as ischemia increase O-GlcNAc levels suggests that other factors also contribute to regulating O-GlcNAc levels. We have previously reported that in unstressed cardiomyocytes exogenous NAD<jats:sup>+<\/jats:sup> resulted in a time and dose dependent decrease in O-GlcNAc levels. Therefore, we postulated that NAD<jats:sup>+<\/jats:sup> and cellular O-GlcNAc levels may be coordinately regulated. Using glucose deprivation as a model system in an immortalized human ventricular cell line, we examined the influence of extracellular NAD<jats:sup>+<\/jats:sup> on cellular O-GlcNAc levels and ER stress in the presence and absence of glucose. We found that NAD<jats:sup>+<\/jats:sup> completely blocked the increase in O-GlcNAc induced by glucose deprivation and suppressed the activation of ER stress. The NAD<jats:sup>+<\/jats:sup> metabolite cyclic ADP-ribose (cADPR) had similar effects on O-GlcNAc and ER stress suggesting a common underlying mechanism. cADPR is a ryanodine receptor (RyR) agonist and like caffeine, which also activates the RyR, both mimicked the effects of NAD<jats:sup>+<\/jats:sup>. SERCA inhibition, which also reduces ER\/SR Ca<jats:sup>2+<\/jats:sup> levels had similar effects to both NAD<jats:sup>+<\/jats:sup> and cADPR on O-GlcNAc and ER stress responses to glucose deprivation. The observation that NAD<jats:sup>+<\/jats:sup>, cADPR, and caffeine all attenuated the increase in O-GlcNAc and ER stress in response to glucose deprivation, suggests a potential common mechanism, linked to ER\/SR Ca<jats:sup>2+<\/jats:sup> levels, underlying their activation. Moreover, we showed that TRPM2, a plasma membrane cation channel was necessary for the cellular responses to glucose deprivation. Collectively, these findings support a novel Ca<jats:sup>2+<\/jats:sup>-dependent mechanism underlying glucose deprivation induced increase in O-GlcNAc and ER stress.<\/jats:p>","DOI":"10.3389\/fmolb.2021.780865","type":"journal-article","created":{"date-parts":[[2021,12,7]],"date-time":"2021-12-07T13:32:41Z","timestamp":1638883961000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":14,"title":["The Identification of a Novel Calcium-Dependent Link Between NAD+ and Glucose Deprivation-Induced Increases in Protein O-GlcNAcylation and ER Stress"],"prefix":"10.3389","volume":"8","author":[{"given":"Luyun","family":"Zou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen E.","family":"Collins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin E.","family":"Young","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhua","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam R.","family":"Wende","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor M.","family":"Darley-Usmar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John C.","family":"Chatham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,12,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1111\/febs.14608","article-title":"Endoplasmic Reticulum Stress Signalling - from Basic Mechanisms to Clinical Applications","volume":"286","author":"Almanza","year":"2019","journal-title":"FEBS J."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1267","DOI":"10.1016\/s0006-291x(02)02073-9","article-title":"ADP-ribose Stimulates the Calcium Release Channel RyR1 in Skeletal Muscle of Rat","volume":"296","author":"Bastide","year":"2002","journal-title":"Biochem. 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