Activation of the Transcriptional Function of the NF-κB Protein c-Rel by O-GlcNAc Glycosylation
Abstract
The transcription factor nuclear factor κB (NF-κB) rapidly reprograms gene expression in response to various stimuli, and its activity is regulated by several posttranslational modifications, including phosphorylation, methylation, and acetylation. The addition of O-linked b-N-acetylglucosamine (a process known as O-GlcNAcylation) is an abundant posttranslational modification that is enhanced in conditions such as hyperglycemia and cellular stress. We report that the NF-κB subunit c-Rel is modified and activated by O-GlcNAcylation. We identified serine 350 as the site of O-GlcNAcylation, which was required for the DNA binding and transactivation functions of c-Rel. Blocking the O-GlcNAcylation of this residue abrogated c-Rel–mediated expression of the cytokine-encoding genes IL2, IFNG, and CSF2 in response to T cell receptor (TCR) activation, whereas increasing the extent of O-GlcNAcylation of cellular proteins enhanced the expression of these genes. TCR- or tumor necrosis factor (TNF)–induced expression of other NF-κB target genes, such as NFKBIA (which encodes IkBa) and TNFAIP3 (which encodes A20), occurred independently of the O-GlcNAcylation of c-Rel. Our findings suggest a stimulus-specific role for hyperglycemia-induced O-GlcNAcylation of c-Rel in promoting T cell–mediated autoimmunity in conditions such as type 1 diabetes by enhancing the production of T helper cell cytokines.
Additional Information
© 2013 American Association for the Advancement of Science. Submitted 22 February 2013; Accepted 9 August 2013; Final Publication 27 August 2013. We express our deep and sincere gratitude to the late N. Sharon for invaluable advice and discussions. We thank A. Weiss for the CD28RE luciferase reporter plasmid; A. Balazs for the lentiviral expression plasmids; the Caltech animal facility; the Caltech protein expression center for baculoviral expression; and J. Rexach, R. Parameswaran, C.-K. Ea, L. Yang, and other members of the Baltimore laboratory for insightful discussions. Funding: This work was initially supported by NIH grant 2R01 GM039458 to D.B., 2RO1 GM084724 to L.C.H.-W., and later by a Mizutani Foundation for Glycoscience grant to P.R. Author contributions: P.R. conceived the project and designed and performed the experiments; P.M.C. performed the PEG labeling; D.E.M. and E.C.P. performed the mass spectrometry; P.R., P.M.C., L.C.H.-W., and D.B. analyzed and interpreted the data; and P.R. and D.B. wrote, and all of the authors edited, the manuscript. Competing interests: A patent was filed for the targeting of c-Rel O-GlcNAcylation (File No. CIT-6437-P).Attached Files
Accepted Version - nihms578080.pdf
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Additional details
- PMCID
- PMC4066889
- Eprint ID
- 41422
- DOI
- 10.1126/scisignal.2004097
- Resolver ID
- CaltechAUTHORS:20130919-142521151
- 2R01 GM039458
- NIH
- 2RO1 GM084724
- NIH
- Mizutani Foundation for Glycoscience
- Created
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2013-09-19Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field