2 219

Cited 35 times in

A dominant negative PPARγ mutant shows altered cofactor recruitment and inhibits adipogenesis in 3T3-L1 cells

Authors
 S. Park  ;  J. L. Jameson  ;  B. D. Freedman  ;  E. J. Lee  ;  Y. Park 
Citation
 DIABETOLOGIA, Vol.46(3) : 365-377, 2003 
Journal Title
 DIABETOLOGIA 
ISSN
 0012-186X 
Issue Date
2003
MeSH
Dominant negative ; PPARγ ; thiazolidinedione ; adipogenesis ; adenovirus ; protein-protein interaction ; 3T3-L1 cells
Keywords
Dominant negative ; PPARγ ; thiazolidinedione ; adipogenesis ; adenovirus ; protein-protein interaction ; 3T3-L1 cells
Abstract
AIMS/HYPOTHESIS: PPARgamma, a member of the nuclear hormone receptor family of transcription factors, plays a key role in adipocyte differentiation and insulin sensitivity. The aim of this study was to identify a potential dominant negative murine PPARgamma mutant and to characterize the in vitro functional properties of this mutant. METHODS: In vitro transient transfections and mammalian two-hybrid assays in TSA201 cells were used to characterize the transcriptional activity of the L466A mutant and to study the molecular interaction of transcriptional cofactors with the L466A mutant in an attempt to elucidate the mechanism of its dominant negative activity. Adenoviral constructs expressing PPARgamma wild-type (AdWT) or the L466A mutant (AdL466A) were infected into the murine 3T3-L1 cell line to study the mutant's effect on adipogenesis. RESULTS: The L466A mutant alone is transcriptionally defective. However, it retains DNA binding and inhibits the ligand-dependent and -independent activity of the wild-type receptor, consistent with dominant negative properties. In mammalian two-hybrid studies, the L466A mutant does not bind nuclear receptor coactivators. However, it more avidly recruits corepressors due to enhanced binding to the corepressor ID1 domain, leading to pronounced transcriptional repression. The AdL466A mutant inhibits adipogenesis induced by either a differentiation cocktail or by thiazolidinedione ligand. AdL466A infection also blocked the upregulation of the adipocyte marker genes aP2 and adipsin. CONCLUSION: We conclude that the L466A PPARgamma mutant possesses potent dominant negative activity based on preferential corepressor recruitment and it inhibits adipogenesis and the expression of adipocyte-specific genes.
Full Text
http://link.springer.com/article/10.1007/s00125-003-1037-4
DOI
10.1007/s00125-003-1037-4
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Lee, Eun Jig(이은직) ORCID logo https://orcid.org/0000-0002-9876-8370
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/113911
사서에게 알리기
  feedback

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse