The transactivating function of peroxisome proliferator-activated receptor gamma is negatively regulated by SUMO conjugation in the amino-terminal domain.
Peroxisome proliferator-activated receptor (PPAR)-gamma2, a member of the nuclear hormone receptor superfamily, plays a key role in adipocyte differentiation. Its amino-terminal region carries a ligand-independent gene-activating function, AF-1, and is composed of activation as well as repression domains. We have found PPARgamma2 and its isoform, PPARgamma1, to be modified by small ubiquitin-related modifier (SUMO)-1 in vivo, at a lysine residue in the repression domain. In reporter assays, a sumoylation-defective K107R mutant of PPARgamma2 exhibited much stronger transactivation than the wild-type, comparable with that of a mutant deleted for the repression domain. A close inverse correlation was observed between the levels of sumoylation and transactivation by PPARgamma2, in analyses employing PPARgamma2 forms with mutations in the sumoylation motif and a dominant-negative mutant of the SUMO conjugating enzyme, Ubc9. Studies with phosphorylation-defective mutants suggested that phosphorylation at S112 of PPARgamma2 promotes K107 sumoylation, and this latter exerts the more potent repressive effects. The K107R mutant PPARgamma2, when infected into NIH3T3 cells with a viral vector, promoted differentiation into adipocytes more efficiently than the wild-type. These observations provide evidence that sumoylation is involved in negative regulation of the transactivating function of PPARgamma2.
Yamashita
Daisuke
D
Graduate School of Life Science, Himeji Institute of Technology, University of Hyogo, 3-2-1 Koto, Kamigori, Hyogo 678-1297, Japan.
Yamaguchi
Tomohiro
T
Shimizu
Makoto
M
Nakata
Nagisa
N
Hirose
Fumiko
F
Osumi
Takashi
T
D006367:HeLa Cells; D006801:Humans; D047495:PPAR gamma / Q000737:chemistry / Q000502:physiology*; D010766:Phosphorylation; D025842:SUMO-1 Protein / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology*
D047495:PPAR gamma; D025842:SUMO-1 Protein