Validation, Identification, and Biological Consequences of the Site-specific O-GlcNAcylation Dynamics of Carbohydrate-responsive Element-binding Protein (ChREBP)*

O-GlcNAcylation of carbohydrate-responsive element-binding protein (ChREBP) is believed as an important modulator of ChREBP activities, however little direct evidence of O-GlcNAcylation on ChREBP and no exact O-GlcNAcylation sites have been reported so far. Here, we validate O-GlcNAcylation on ChREBP in cell-free coupled transcription/translation system and in cells by chemoenzymatic and metabolic labeling, respectively. Moreover, for the first time, we identify O-GlcNAcylation on Ser614 in the C-terminus of ChREBP by mass spectrometry and validate two important sites, Thr517 and Ser839 for O-GlcNAcylation and their function via molecular and chemical biological method. Under high glucose conditions, Ser514 phosphorylation enhances ChREBP O-GlcNAcylation, maintaining the transcriptional activity of ChREBP; Ser839 O-GlcNAcylation is essential for Mlx-heterodimerization and DNA-binding activity enhancement, consequently inducing transcriptional activity. Ser839 O-GlcNAcylation is also crucial for ChREBP nuclear export partially by strengthening interactions with CRM1 and 14-3-3. This work is a detailed study of ChREBP O-GlcNAcylation and highlights the biological consequences of the site-specific O-GlcNAcylation dynamics of ChREBP.

• Publication year

  2017

• Venue

  Molecular & Cellular Proteomics

• Publication date

  2017-04-27

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/mcp.M116.061416PMID 28450420PMCID PMC5500757
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• 2016 update of the PRIDE database and its related tools

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  2012cited by this paper
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  2012cited by this paper
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  2012cited by this paper
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  2012cited by this paper
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  2011cited by this paper
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  2011cited by this paper
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  2011cited by this paper
• Phosphoproteomic Profiling of In Vivo Signaling in Liver by the Mammalian Target of Rapamycin Complex 1 (mTORC1)

  2011influential reference
• Combining high-energy C-trap dissociation and electron transfer dissociation for protein O-GlcNAc modification site assignment.

  2011cited by this paper
• Salt-inducible kinase 2 links transcriptional coactivator p300 phosphorylation to the prevention of ChREBP-dependent hepatic steatosis in mice.

  2010cited by this paper
• Quantitation of O-Glycosylation Stoichiometry and Dynamics using Resolvable Mass Tags

  2010cited by this paper
• Coordinate regulation/localization of the carbohydrate responsive binding protein (ChREBP) by two nuclear export signal sites: discovery of a new leucine-rich nuclear export signal site.

  2010cited by this paper
• The role of O-linked GlcNAc modification on the glucose response of ChREBP.

  2010cited by this paper
• The glucose-responsive transcription factor ChREBP contributes to glucose-dependent anabolic synthesis and cell proliferation

  2009cited by this paper
• Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins.

  2008cited by this paper
• Identification and function of phosphorylation in the glucose-regulated transcription factor ChREBP.

  2008influential reference
• A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo.

  2008influential reference
• Regulation of Nuclear Import/Export of Carbohydrate Response Element-binding Protein (ChREBP)

  2008cited by this paper
• Glucose Is Necessary for Embryonic Pancreatic Endocrine Cell Differentiation*

  2007cited by this paper
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  2006cited by this paper
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  2006cited by this paper
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  2006cited by this paper
• Deficiency of carbohydrate-activated transcription factor ChREBP prevents obesity and improves plasma glucose control in leptin-deficient (ob/ob) mice.

  2006cited by this paper
• Metabolic syndrome.

  2005cited by this paper
• Deficiency of carbohydrate response element-binding protein (ChREBP) reduces lipogenesis as well as glycolysis.

  2004influential reference
• Mlx Is the Functional Heteromeric Partner of the Carbohydrate Response Element-binding Protein in Glucose Regulation of Lipogenic Enzyme Genes*

  2004cited by this paper
• Carbohydrate response element binding protein directly promotes lipogenic enzyme gene transcription.

  2004influential reference
• The subcellular localization of the ChoRE-binding protein, encoded by the Williams-Beuren syndrome critical region gene 14, is regulated by 14-3-3.

  2004cited by this paper
• The metabolic syndrome.

  2003cited by this paper
• A Novel Heterodimerization Domain, CRM1, and 14-3-3 Control Subcellular Localization of the MondoA-Mlx Heterocomplex

  2002cited by this paper
• Glucose and cAMP regulate the L-type pyruvate kinase gene by phosphorylation/dephosphorylation of the carbohydrate response element binding protein

  2001cited by this paper
• A glucose-responsive transcription factor that regulates carbohydrate metabolism in the liver

  2001influential reference
• WBSCR14, a gene mapping to the Williams--Beuren syndrome deleted region, is a new member of the Mlx transcription factor network.

  2001influential reference
• MondoA, a Novel Basic Helix-Loop-Helix–Leucine Zipper Transcriptional Activator That Constitutes a Positive Branch of a Max-Like Network

  2000cited by this paper
• Mechanisms by which carbohydrates regulate expression of genes for glycolytic and lipogenic enzymes.

  1997cited by this paper
• Two CACGTG Motifs with Proper Spacing Dictate the Carbohydrate Regulation of Hepatic Gene Transcription (*)

  1995cited by this paper
• Purification and characterization of an O-GlcNAc selective N-acetyl-beta-D-glucosaminidase from rat spleen cytosol.

  1994cited by this paper
• Glycosylation of nuclear and cytoplasmic proteins. Purification and characterization of a uridine diphospho-N-acetylglucosamine:polypeptide beta-N-acetylglucosaminyltransferase.

  1992cited by this paper
• Glucose stimulation of lipogenic enzyme gene expression in cultured white adipose tissue. A role for glucose 6-phosphate.

  1992cited by this paper
• Glycosylation of nuclear pore protein p62. Reticulocyte lysate catalyzes O-linked N-acetylglucosamine addition in vitro.

  1990cited by this paper
• O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation.

  1988cited by this paper

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  2025cites this paper
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• A high throughput screen for pharmacological inhibitors of the carbohydrate response element

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  2021cites this paper
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  2021cites this paper
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  2021cites this paper
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  2020cites this paper
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  2020cites this paper
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  2020cites this paper
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  2019cites this paper
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  2019cites this paper
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  2019cites this paper
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