title: Modification of RelA by O-linked N-acetylglucosamine links glucose metabolism to NF-κB acetylation and transcription.
abstract: The molecular mechanisms linking glucose metabolism with active transcription remain undercharacterized in mammalian cells. Using nuclear factor-κB (NF-κB) as a glucose-responsive transcription factor, we show that cells use the hexosamine biosynthesis pathway and O-linked β-N-acetylglucosamine (O-GlcNAc) transferase (OGT) to potentiate gene expression in response to tumor necrosis factor (TNF) or etoposide. Chromatin immunoprecipitation assays demonstrate that, upon induction, OGT localizes to NF-κB-regulated promoters to enhance RelA acetylation. Knockdown of OGT abolishes p300-mediated acetylation of RelA on K310, a posttranslational mark required for full NF-κB transcription. Mapping studies reveal T305 as an important residue required for attachment of the O-GlcNAc moiety on RelA. Furthermore, p300 fails to acetylate a full-length RelA(T305A) mutant, linking O-GlcNAc and acetylation events on NF-κB. Reconstitution of RelA null cells with the RelA(T305A) mutant illustrates the importance of this residue for NF-κB-dependent gene expression and cell survival. Our work provides evidence for a unique regulation where attachment of the O-GlcNAc moiety to RelA potentiates p300 acetylation and NF-κB transcription.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22908, USA.
authors.forename: David F
authors.initials: DF
authors.lastname: Allison
authors.forename: J Jacob
authors.initials: JJ
authors.lastname: Wamsley
authors.forename: Manish
authors.initials: M
authors.lastname: Kumar
authors.forename: Duo
authors.initials: D
authors.lastname: Li
authors.forename: Lisa G
authors.initials: LG
authors.lastname: Gray
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Hart
authors.forename: David R
authors.initials: DR
authors.lastname: Jones
authors.forename: Marty W
authors.initials: MW
authors.lastname: Mayo
mesh_terms: D000107:Acetylation; D000117:Acetylglucosamine / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005047:Etoposide / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D005947:Glucose / Q000378:metabolism*; D057809:HEK293 Cells; D006595:Hexosamines / Q000096:biosynthesis; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008156:Luciferases; D053858:Metabolic Networks and Pathways / Q000235:genetics / Q000502:physiology*; D016328:NF-kappa B / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D006595:Hexosamines; D016328:NF-kappa B; C498143:RELA protein, human; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D005047:Etoposide; D008156:Luciferases; D005947:Glucose; D000117:Acetylglucosamine