A SUMOylation-dependent pathway mediates transrepression of inflammatory response genes by PPAR-γ

Abstract

Peroxisome proliferator-activated receptor-γ (PPAR-γ) has essential roles in adipogenesis and glucose homeostasis, and is a molecular target of insulin-sensitizing drugs1,2,3. Although the ability of PPAR-γ agonists to antagonize inflammatory responses by transrepression of nuclear factor kappa B (NF-κB) target genes is linked to antidiabetic4 and antiatherogenic actions5, the mechanisms remain poorly understood. Here we report the identification of a molecular pathway by which PPAR-γ represses the transcriptional activation of inflammatory response genes in mouse macrophages. The initial step of this pathway involves ligand-dependent SUMOylation of the PPAR-γ ligand-binding domain, which targets PPAR-γ to nuclear receptor corepressor (NCoR)–histone deacetylase-3 (HDAC3) complexes on inflammatory gene promoters. This in turn prevents recruitment of the ubiquitylation/19S proteosome machinery that normally mediates the signal-dependent removal of corepressor complexes required for gene activation. As a result, NCoR complexes are not cleared from the promoter and target genes are maintained in a repressed state. This mechanism provides an explanation for how an agonist-bound nuclear receptor can be converted from an activator of transcription to a promoter-specific repressor of NF-κB target genes that regulate immunity and homeostasis.

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Figure 1: PPAR-γ prevents LPS-induced dissociation of the NCoR–HDAC3 complex from the iNOS promoter.
Figure 2: PIAS1 interacts with PPAR-γ and is required for transrepression of iNOS.
Figure 3: Ligand-dependent SUMOylation of PPAR-γ is required for transrepression.
Figure 4: SUMOylation of PPAR-γ promotes interaction with the NCoR–HDAC3 complex.

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Acknowledgements

We thank A. Beas, W. Books and J. Goode for assistance with the yeast two-hybrid screen, and J. Collier for assistance with the mammalian two-hybrid assays. We thank F. Almazan for RAW264.7 cells. We thank M. Ricote for helpful discussions and M. Lambert for assistance with analysis of PPAR-γ crystal structures. We thank A. Howarth for assistance with preparation of the manuscript. These studies were supported by an American Heart Association Predoctoral Fellowship to G.P. and National Institutes of Health grants to D.W.R., M.G.R. and C.K.G, and support from the Stanford Donald W. Reynolds Cardiovascular Center to C.K.G. M.G.R. is supported by the Howard Hughes Medical Institute. Author Contributions G.P., A.L.F., S.O., A.G., A.C.L. and D.W.R. performed experimental work and data analysis. T.M.W., M.G.R. and C.K.G. performed data analysis. C.K.G. was responsible for project planning.

Author information

Correspondence to Christopher K. Glass.

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Competing interests

T.M.W. is an employee of GlaxoSmithKline.

Supplementary information

Supplementary Figure S1

Efficacy of siRNAs directed against NCoR, HDAC3, HDAC7, PIAS1 and Ubc9. (PDF 112 kb)

Supplementary Figure S2

NCoR and HDAC3 are required for PPARγ-mediated transrepression. (PDF 115 kb)

Supplementary Figure S3

PPARγ transrepression does not require sequence specific DNA binding and can be extended to other LPS-dependent promoters. (PDF 163 kb)

Supplementary Figure S4

PIAS1 interacts with PPARγ and is required for ligand-dependent inhibition of iNOS gene activation. Supplementary Figure S4 was replaced on 03 February 2006. This figure replaces the erroneous posting of Figure 4 from the main text as Supplementary Figure 4. (PDF 534 kb)

Supplementary Figure S5

Sumoylation is required for PPARγ transrepression and enhances ligand dependent association with NCoR. (PDF 132 kb)

Supplementary Figure Legends

Text to accompany the above Supplementary Figures. (DOC 23 kb)

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