Regulation of nuclear import/export of carbohydrate response element-binding protein (ChREBP): Interaction of anα-helix of ChREBP with the 14-3-3 proteins and regulation by phosphorylation

Haruhiko Sakiyama, R. Max Wynn, Wan Ru Lee, Masashi Fukasawa, Hiroyuki Mizuguchi, Kevin H. Gardner, Joyce J. Repa, Kosaku Uyeda

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

Carbohydrate response element-binding protein (ChREBP) is a glucose-responsive transcription factor that plays a critical role in the glucose-mediated induction of gene products involved in hepatic glycolysis and lipogenesis. Glucose affects the activity of ChREBP largely through post-translational mechanisms involving phosphorylation-dependent cellular localization. In this work we show that the N-terminal region of ChREBP (residues 1-251) regulates its subcellular localization via an interaction with 14-3-3. 14-3-3 binds an α-helix in this region (residues 125-135) to retain ChREBP in the cytosol, and binding of 14-3-3 is facilitated by phosphorylation of nearby Ser-140 and Ser-196. Phosphorylation of ChREBP at these sites was essential for its interaction with CRM1 for export to the cytosol, whereas nuclear import of ChREBP requires dephosphorylated ChREBP to interact with importin α. Notably, 14-3-3 appears to compete with importin α for ChREBP binding. 14-3-3β bound to a synthetic peptide spanning residues 125-144 and bearing a phosphate at Ser-140 with a dissociation constant of 1.1 μM, as determined by isothermal calorimetry. The interaction caused a shift in the fluorescence maximum of the tryptophan residues of the peptide. The corresponding unphosphorylated peptide failed to bind 14-3-3β. These results suggest that interactions with importin α and 14-3-3 regulate movement of ChREBP into and out of the nucleus, respectively, and that these interactions are regulated by the ChREBP phosphorylation status.

Original languageEnglish (US)
Pages (from-to)24899-24908
Number of pages10
JournalJournal of Biological Chemistry
Volume283
Issue number36
DOIs
StatePublished - Sep 5 2008

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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