TATA-Box Binding Protein O-GlcNAcylation at T114 Regulates Formation of the B-TFIID Complex and Is Critical for Metabolic Gene Regulation

Mol Cell. 2020 Mar 5;77(5):1143-1152.e7. doi: 10.1016/j.molcel.2019.11.022. Epub 2019 Dec 19.

Abstract

In eukaryotes, gene expression is performed by three RNA polymerases that are targeted to promoters by molecular complexes. A unique common factor, the TATA-box binding protein (TBP), is thought to serve as a platform to assemble pre-initiation complexes competent for transcription. Here, we describe a novel molecular mechanism of nutrient regulation of gene transcription by dynamic O-GlcNAcylation of TBP. We show that O-GlcNAcylation at T114 of TBP blocks its interaction with BTAF1, hence the formation of the B-TFIID complex, and its dynamic cycling on and off of DNA. Transcriptomic and metabolomic analyses of TBPT114A CRISPR/Cas9-edited cells showed that loss of O-GlcNAcylation at T114 increases TBP binding to BTAF1 and directly impacts expression of 408 genes. Lack of O-GlcNAcylation at T114 is associated with a striking reprogramming of cellular metabolism induced by a profound modification of the transcriptome, leading to gross alterations in lipid storage.

Keywords: B-TFIID; BTAF1; O-GlcNAc; O-GlcNAcylation; TATA-box binding protein; TBP; lipid droplets; metabolism; nutrient sensing; transcription.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / genetics
  • Chromatin / metabolism
  • Diabetes Mellitus, Experimental / genetics
  • Diabetes Mellitus, Experimental / metabolism
  • Gene Expression Regulation
  • Glucose / metabolism*
  • Glycosylation
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Lipid Droplets / metabolism*
  • Lipid Metabolism* / genetics
  • Male
  • Multiprotein Complexes
  • Rats, Sprague-Dawley
  • Signal Transduction
  • TATA-Binding Protein Associated Factors / genetics
  • TATA-Binding Protein Associated Factors / metabolism*
  • TATA-Box Binding Protein / genetics
  • TATA-Box Binding Protein / metabolism*
  • Time Factors
  • Transcription Factor TFIID / genetics
  • Transcription Factor TFIID / metabolism*
  • Transcription, Genetic
  • Transcriptome

Substances

  • BTAF1 protein, human
  • Chromatin
  • Multiprotein Complexes
  • TATA-Binding Protein Associated Factors
  • TATA-Box Binding Protein
  • TBP protein, human
  • Transcription Factor TFIID
  • Glucose