RORγ is a nuclear receptor that binds to DNA motifs as a monomer to constitutively activate target genes. RORγ plays an important role in thymocyte development and lymph node organogenesis, while the regulation of RORγ-mediated transcriptional activation is currently unclear. The purpose of this study was to identify other nuclear proteins that interact with RORγ. A yeast two-hybrid screen with Y190 yeast cells under stringent conditions resulted in the identification of CHD4, also known as Mi-2β, as a RORγ-interacting protein. This interaction was confirmed by GST pull-down assays. This interaction occurred within the middle regulatory region (amino acids 719-1164) of Mi-2β. Transfection of Gal4-RORγ into HeLa cells resulted in constitutive transactivation of the MH100-tk-luc reporter. The addition of Mi-2β resulted in a dramatic 50% decrease in Gal4-RORγ-mediated transactivation. These data demonstrate that RORγ forms a protein-protein interaction with the regulatory region of Mi-2β, resulting in inhibition of RORγ transcriptional activity. These results may provide evidence as to how RORγ-mediated transactivation is regulated by other nuclear proteins.
|Original language||English (US)|
|Number of pages||5|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Jun 4 2004|
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology