TY - JOUR
T1 - PPARγ lipodystrophy mutants reveal intermolecular interactions required for enhancer activation
AU - Madsen, Maria Stahl
AU - Broekema, Marjoleine F.
AU - Madsen, Martin Rønn
AU - Koppen, Arjen
AU - Borgman, Anouska
AU - Gräwe, Cathrin
AU - Thomsen, Elisabeth G.K.
AU - Westland, Denise
AU - Kranendonk, Mariette E.G.
AU - Koerkamp, Marian Groot
AU - Hamers, Nicole
AU - Bonvin, Alexandre M.J.J.
AU - Pittol, José M.Ramos
AU - Natarajan, Kedar Nath
AU - Kersten, Sander
AU - Holstege, Frank C.P.
AU - Monajemi, Houshang
AU - van Mil, Saskia W.C.
AU - Vermeulen, Michiel
AU - Kragelund, Birthe B.
AU - Cassiman, David
AU - Mandrup, Susanne
AU - Kalkhoven, Eric
N1 - © 2022. The Author(s).
PY - 2022/11/19
Y1 - 2022/11/19
N2 - 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.
AB - 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.
KW - Humans
KW - PPAR gamma/genetics
KW - Adipocytes/metabolism
KW - Retinoid X Receptors/genetics
KW - Lipodystrophy/metabolism
KW - Regulatory Sequences, Nucleic Acid
UR - http://www.scopus.com/inward/record.url?scp=85142129834&partnerID=8YFLogxK
U2 - 10.1038/s41467-022-34766-9
DO - 10.1038/s41467-022-34766-9
M3 - Article
C2 - 36402763
AN - SCOPUS:85142129834
SN - 2041-1723
VL - 13
SP - 7090
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 7090
ER -