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Hepatocyte nuclear factor 1 alpha (HNF1A) regulates transcription of O‐GlcNAc transferase in a negative feedback mechanism

O‐GlcNAc transferase (OGT)‐catalyzed protein O‐GlcNAcylation is implicated in diverse cellular events. In the present study, we report the regulation of ogt transcription by the hepatocyte nuclear factor 1 homologue A (HNF1A) in HEK293T cells. We first identified a core ogt promoter (−150 to +200 bp) and confirmed its binding to the transcription factor HNF1A. We found that HNF1A regulates ogt transcription in a time‐dependent manner and that O‐GlcNAcylation of HNF1A represses ogt transcription. Electron‐transfer dissociation based tandem mass spectrometry analysis revealed 14 O‐GlcNAc sites on HNF1A, six of which are predominantly modified, including Ser303/304, Ser471, Ser560 and Thr563/564. We further found that loss of O‐GlcNAcylation at Ser303/304 or Thr563/564 significantly elevates ogt transcription. These findings highlight a negative feedback mechanism for ogt transcription, which partially explains the homeostasis of cellular O‐GlcNAcylation.