O-linked N-acetylglucosamine modification on CCAAT enhancer-binding protein β. Role during adipocyte differentiation

Xi Li, Henrik Molina, Haiyan Huang, You You Zhang, Mei Liu, Shu Wen Qian, Chad Slawson, Wagner B. Dias, Akhilesh Pandey, Gerald W. Hart, M. Daniel Lane, Qi Qun Tang

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

CCAAT enhancer-binding protein (C/EBP)β is a basic leucine zipper transcription factor family member, and can be phosphorylated, acetylated, and sumoylated. C/EBPβ undergoes sequential phosphorylation during 3T3-L1 adipocyte differentiation. Phosphorylation on Thr188 by MAPK or cyclin A/cdk2 primes the phosphorylations on Ser184/Thr179 by GSK3β, and these phosphorylations are required for the acquisition of DNA binding activity of C/EBPβ. Here we show that C/EBPβ is modified by O-GlcNAc, a dynamic single sugar modification found on nucleocytoplasmic proteins. The GlcNAcylation sites are Ser180 and Ser181, which are in the regulation domain and are very close to the phosphorylation sites (Thr188, Ser184, and Thr179) required for the gain of DNA binding activity. Both in vitro and ex vivo experiments demonstrate that GlcNAcylation on Ser180 and Ser181 prevents phosphorylation on Thr188, Ser184, and Thr179, as indicated by the decreased relative phosphorylation and DNA binding activity of C/EBPβ delayed the adipocyte differentiation program. Mutation of both Ser180 and Ser181 to Ala significantly increase the transcriptional activity of C/EBPβ. These data suggest that GlcNAcylation regulates both the phosphorylation and DNA binding activity of C/EBPβ.

Original languageEnglish (US)
Pages (from-to)19248-19254
Number of pages7
JournalJournal of Biological Chemistry
Volume284
Issue number29
DOIs
StatePublished - Jul 17 2009

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Li, X., Molina, H., Huang, H., Zhang, Y. Y., Liu, M., Qian, S. W., Slawson, C., Dias, W. B., Pandey, A., Hart, G. W., Lane, M. D., & Tang, Q. Q. (2009). O-linked N-acetylglucosamine modification on CCAAT enhancer-binding protein β. Role during adipocyte differentiation. Journal of Biological Chemistry, 284(29), 19248-19254. https://doi.org/10.1074/jbc.M109.005678