title: Hsp90 regulates O-linked β-N-acetylglucosamine transferase: a novel mechanism of modulation of protein O-linked β-N-acetylglucosamine modification in endothelial cells.
abstract: O-linked β-N-acetylglucosamine (O-GlcNAc) modification of proteins is involved in many important cellular processes. Increased O-GlcNAc has been implicated in major diseases, such as diabetes and its complications and cardiovascular and neurodegenerative diseases. Recently, we reported that O-GlcNAc modification occurs in the proteasome and serves to inhibit proteasome function by blocking the ATPase activity in the 19S regulatory cap, explaining, at least in part, the adverse effects of O-GlcNAc modification and suggesting that downregulating O-GlcNAc might be important in the treatment of human diseases. In this study, we report on a novel mechanism to modulate cellular O-GlcNAc modification, namely through heat shock protein 90 (Hsp90) inhibition. We observed that O-linked β-N-acetylglucosamine transferase (OGT) interacts with the tetratricopeptide repeat binding site of Hsp90. Inhibition of Hsp90 by its specific inhibitors, radicicol or 17-N-allylamino-17-demethoxygeldanamycin, destabilized OGT in primary endothelial cell cultures and enhanced its degradation by the proteasome. Furthermore, Hsp90 inhibition downregulated O-GlcNAc protein modifications and attenuated the high glucose-induced increase in O-GlcNAc protein modification, including high glucose-induced increase in endothelial or type 3 isoform of nitric oxide synthase (eNOS) O-GlcNAcylation. These results suggest that Hsp90 is involved in the regulation of OGT and O-GlcNAc modification and that Hsp90 inhibitors might be used to modulate O-GlcNAc modification and reverse its adverse effects in human diseases.
authors.affiliation: Vascular Biology Center, Medical College of Georgia, Georgia Health Sciences University, Augusta, 30912-2500, USA. fzhang@georgiahealth.edu
authors.forename: Fengxue
authors.initials: F
authors.lastname: Zhang
authors.forename: Connie M
authors.initials: CM
authors.lastname: Snead
authors.forename: John D
authors.initials: JD
authors.lastname: Catravas
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology; D001665:Binding Sites; D002417:Cattle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D004795:Enzyme Stability; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D018942:Macrolides / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D013997:Time Factors; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics
chemical_list: D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D018942:Macrolides; C112765:tanespimycin; D052250:Nitric Oxide Synthase Type III; C492951:Stub1 protein, mouse; D044767:Ubiquitin-Protein Ligases; D017351:N-Acetylglucosaminyltransferases; C063057:UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase; D046988:Proteasome Endopeptidase Complex; C035359:monorden; D005947:Glucose; D000117:Acetylglucosamine