PPARγ lipodystrophy mutants reveal intermolecular interactions required for enhancer activation

Maria Stahl Madsen, Marjoleine F. Broekema, Martin Rønn Madsen, Arjen Koppen, Anouska Borgman, Cathrin Gräwe, Elisabeth G.K. Thomsen, Denise Westland, Mariette E.G. Kranendonk, Marian Groot Koerkamp, Nicole Hamers, Alexandre M.J.J. Bonvin, José M.Ramos Pittol, Kedar Nath Natarajan, Sander Kersten, Frank C.P. Holstege, Houshang Monajemi, Saskia W.C. van Mil, Michiel Vermeulen, Birthe B. KragelundDavid Cassiman, Susanne Mandrup*, Eric Kalkhoven


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Peroxisome proliferator-activated receptor γ (PPARγ) is the master regulator of adipocyte differentiation, and mutations that interfere with its function cause lipodystrophy. PPARγ is a highly modular protein, and structural studies indicate that PPARγ domains engage in several intra- and inter-molecular interactions. How these interactions modulate PPARγ’s ability to activate target genes in a cellular context is currently poorly understood. Here we take advantage of two previously uncharacterized lipodystrophy mutations, R212Q and E379K, that are predicted to interfere with the interaction of the hinge of PPARγ with DNA and with the interaction of PPARγ ligand binding domain (LBD) with the DNA-binding domain (DBD) of the retinoid X receptor, respectively. Using biochemical and genome-wide approaches we show that these mutations impair PPARγ function on an overlapping subset of target enhancers. The hinge region-DNA interaction appears mostly important for binding and remodelling of target enhancers in inaccessible chromatin, whereas the PPARγ-LBD:RXR-DBD interface stabilizes the PPARγ:RXR:DNA ternary complex. Our data demonstrate how in-depth analyses of lipodystrophy mutants can unravel molecular mechanisms of PPARγ function.

TidsskriftNature Communications
Udgave nummer1
StatusUdgivet - dec. 2022

Bibliografisk note

Funding Information:
This work was supported by grants from the Independent Research Fund Denmark (Sapere Aude Advanced grant no. 12-125524 to S.M.), the Danish National Research Foundation (DNRF grant no. 141 to the Center for Functional Genomics and Tissue Plasticity (ATLAS) (to S.M.)), the Novo Nordisk Foundation (support to S.M. through the NNF Center for Stem Cell Biology (NNF17CC0027852) and to BBK through the NNF Challenge center REPIN (NNF18OC0033926) and cOpenNMR (NNF18OC0032996), (the Villum Foundation (support to S.M. through the Villum Center for Bioanalytical Sciences and to K.N.N through grant VYI25397). The Vermeulen lab is part of the Oncode Institute, which is partly funded by the Dutch Cancer Society. We thank members of the Kalkhoven, Mandrup and Van Mil laboratories for helpful discussions and Dr. Wilko Spiering for clinical assessment. We thank Dr. Natasja Rochel and Dr. Bruno Klaholz for kindly providing the SAXS data of the DNA-bound PPARγ/RXRα heterodimer structure, Marijke Baltissen and Dr. Cornelia G. Spruijt for expert technical advice, Dr. Johan de Rooij for the pLV-CMV-FLAG-betaCatenin-Ires-PURO vector, and Dr. A. Bensadoun for kindly providing the antibody against mouse LPL.

Publisher Copyright:
© 2022, The Author(s).


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