Structural characterization of a unique interface between carbohydrate response element-binding protein (ChREBP) and 14-3-3β protein

Qiang Ge, Nian Huang, R. Max Wynn, Yang Li, Xinlin Du, Bonnie Miller, Hong Zhang, Kosaku Uyeda

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Carbohydrate response element-binding protein (ChREBP) is an insulin-independent, glucose-responsive transcription factor that is expressed at high levels in liver hepatocytes where it plays a critical role in converting excess carbohydrates to fat for storage. In response to fluctuating glucose levels, hepatic ChREBP activity is regulated in large part by nucleocytoplasmic shuttling of ChREBP protein via interactions with 14-3-3 proteins. The N-terminal ChREBP regulatory region is necessary and sufficient for glucose-responsive ChREBP nuclear import and export. Here, we report the crystal structure of a complex of 14-3-3β bound to the N-terminal regulatory region of ChREBP at 2.4 Å resolution. The crystal structure revealed that the α2 helix of ChREBP (residues 117-137) adopts a well defined α-helical conformation and binds 14-3-3 in a phosphorylation-independent manner that is different from all previously characterized 14-3-3 and target protein-binding modes. ChREBP α2 interacts with 14-3-3 through both electrostatic and van der Waals interactions, and the binding is partially mediated by a free sulfate or phosphate. Structure-based mutagenesis and binding assays indicated that disrupting the observed 14-3-3 and ChREBP α2 interface resulted in a loss of complex formation, thus validating the novel protein interaction mode in the 14-3-3β•ChREBP α2 complex.

Original languageEnglish (US)
Pages (from-to)41914-41921
Number of pages8
JournalJournal of Biological Chemistry
Volume287
Issue number50
DOIs
StatePublished - Dec 7 2012

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14-3-3 Proteins
Response Elements
Carrier Proteins
Carbohydrates
Cell Nucleus Active Transport
Nucleic Acid Regulatory Sequences
Glucose
Crystal structure
Mutagenesis
Phosphorylation
Liver
Static Electricity
Protein Binding
Sulfates
Conformations
Hepatocytes
Electrostatics
Assays
Proteins
Transcription Factors

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Structural characterization of a unique interface between carbohydrate response element-binding protein (ChREBP) and 14-3-3β protein. / Ge, Qiang; Huang, Nian; Wynn, R. Max; Li, Yang; Du, Xinlin; Miller, Bonnie; Zhang, Hong; Uyeda, Kosaku.

In: Journal of Biological Chemistry, Vol. 287, No. 50, 07.12.2012, p. 41914-41921.

Research output: Contribution to journalArticle

Ge, Qiang ; Huang, Nian ; Wynn, R. Max ; Li, Yang ; Du, Xinlin ; Miller, Bonnie ; Zhang, Hong ; Uyeda, Kosaku. / Structural characterization of a unique interface between carbohydrate response element-binding protein (ChREBP) and 14-3-3β protein. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 50. pp. 41914-41921.
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