Genome-wide analysis of E157D PPARγ transcriptional activity in NIH-3T3 fibroblasts

The E157D PPARγ mutation presents as an atypical case of familial partial lipodystrophy and diabetes in a human cohort and has a novel molecular mechanism. The mutation is located in the DNA-binding domain, has no dominant negative activity but disrupts transcription activation after the protein has bound the DNA. The mutation causes a transcriptional activation defect on many PPARγ target genes, while some genes are induced normally. A small number of non-PPARγ target genes are induced by the E157D mutant, possibly leading to the atypical features in this cohort. mRNA was amplified from total RNA collected from NIH-3T3 cells expressing either wild-type or E157D PPARγ through lentiviral infection. Cells were treated with DMSO or 20µM rosiglitazone for 24 hours prior to RNA isolation in three independent experiments.