Direct association between the CREB-binding protein (CBP) and nuclear receptor corepressor (N-CoR).
The binding of ligand to nuclear hormone receptors induces a conformational change that results in corepressor release and the recruitment of coactivator proteins that contain or recruit histone acetyltransferase (HAT) activity. As such, the coactivator and corepressor complexes and their associated HAT and histone deacytlase (HDAC) activities are often believed to be segregated into distinct complexes. However, there have been several reports that suggest that coactivators and corepressors may not be strictly segregated and in some cases even interact directly. In the present study, we have utilized a biochemical approach to assess whether the nuclear receptor corepressor (N-CoR) is capable of associating with the HAT coactivator CREB-binding protein (CBP). We demonstrate, using both immunoaffinity purification and conventional chromatography, that a subset of the N-CoR-HDAC3 complex copurifies with CBP in HeLa cells. In addition, indirect immunofluorescence also indicates an association between N-CoR and CBP in intact MCF-7 cells. This association may be direct as in vitro pulldown assays using recombinant purified proteins indicated that the amino terminus of N-CoR interacts directly with CBP. Interestingly, we also demonstrate that increasing concentrations of N-CoR are capable of attenuating CBP HAT activity in vitro, suggesting that N-CoR may have a functional role in modulating HAT activity. This is the first report of a direct interaction between N-CoR and CBP, and suggests that the role of N-CoR in mediating transcriptional events may be more complex than previously anticipated.
Cowger
Jeffery John Michael
JJ
Department of Oncology, London Regional Cancer Program, The University of Western Ontario, London, Ontario N6A 4L6, Canada.
Torchia
Joseph
J
D015153:Blotting, Western; D050882:CREB-Binding Protein / Q000378:metabolism*; D002846:Chromatography, Affinity; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D009687:Nuclear Proteins / Q000378:metabolism*; D056971:Nuclear Receptor Co-Repressor 1; D012097:Repressor Proteins / Q000378:metabolism*
C539127:NCOR1 protein, human; D009687:Nuclear Proteins; D056971:Nuclear Receptor Co-Repressor 1; D012097:Repressor Proteins; D050882:CREB-Binding Protein