Protein-DNA array-based identification of transcription factor activities regulated by interaction with the glucocorticoid receptor.
The glucocorticoid receptor (GR) regulates gene expression by binding specific sequence elements within the promoters of target genes or by cross-talk with other transcription factors (TFs). For some TFs, interaction with the GR results in alteration of DNA binding and transcriptional regulation. We used a protein-DNA array, a system that facilitates simultaneous profiling of the activities of multiple transcription factors, to systematically examine the potential cross-talk of GRalpha with 149 TFs. Using this array, we identified several TFs, including IRF, E47, and COUP-TF, whose DNA binding activities were modulated by GRalpha. We then confirmed these results with in vitro electrophoretic mobility shift assays and in vivo reporter assays. In this study, IRF and E47 were identified as participants in GRalpha cross-talk for the first time. This new finding expands our understanding of the functional role of GRalpha in the context of gene expression regulation.
Jiang
Xin
X
Panomics, Inc., Redwood City, California 94063 and LINE, Bristol University, Bristol BS1 3NY, United Kingdom. xjiang@panomics.com
Norman
Michael
M
Roth
Leslie
L
Li
Xianqiang
X
D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D051836:COUP Transcription Factors; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D050856:Interferon Regulatory Factor-1; D008156:Luciferases / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010750:Phosphoproteins / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051820:TCF Transcription Factors; D058125:Transcription Factor 7-Like 1 Protein; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection
D000199:Actins; D051836:COUP Transcription Factors; D004268:DNA-Binding Proteins; C494257:IRF1 protein, human; D050856:Interferon Regulatory Factor-1; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D051820:TCF Transcription Factors; C544912:TCF7L1 protein, human; D058125:Transcription Factor 7-Like 1 Protein; D014157:Transcription Factors; D012313:RNA; D004247:DNA; D008156:Luciferases