EP300-Mediated Lysine 2-Hydroxyisobutyrylation Regulates Glycolysis

Lysine 2-hydroxyisobutyrylation (Khib) is an evolutionarily conserved and widespread histone mark like lysine acetylation (Kac). Here we report that EP300 functions as a lysine 2-hyroxyisobutyryltransferase to regulate glycolysis in response to nutritional cues. We discovered that EP300 differentially regulates Khib and Kac on distinct lysine sites, with only 6 of the 149 EP300-targeted Khib sites overlapping with the 693 EP300-targeted Kac sites. We demonstrate that diverse cellular proteins, particularly glycolytic enzymes, are targeted by EP300 for Khib, but not for Kac. Specifically, deletion of EP300 significantly reduces Khib levels on several EP300-dependent, Khib-specific sites on key glycolytic enzymes including ENO1, decreasing their catalytic activities. Consequently, EP300-deficient cells have impaired glycolysis and are hypersensitive to glucose-depletion-induced cell death. Our study reveals an EP300-catalyzed, Khib-specific molecular mechanism that regulates cellular glucose metabolism and further indicate that EP300 has an intrinsic ability to select short-chain acyl-CoA-dependent protein substrates.