Protein-DNA array-based identification of transcription factor activities regulated by interaction with the glucocorticoid receptor

Xin Jiang; Michael Norman; Leslie Roth; Xianqiang Li

doi:10.1074/jbc.M403948200

Protein-DNA array-based identification of transcription factor activities regulated by interaction with the glucocorticoid receptor

J Biol Chem. 2004 Sep 10;279(37):38480-5. doi: 10.1074/jbc.M403948200. Epub 2004 Jul 7.

Authors

Xin Jiang¹, Michael Norman, Leslie Roth, Xianqiang Li

Affiliation

¹ Panomics, Inc., Redwood City, California 94063 and LINE, Bristol University, Bristol BS1 3NY, United Kingdom. [email protected]

PMID: 15247296
DOI: 10.1074/jbc.M403948200

Abstract

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.

MeSH terms

Actins / metabolism
Animals
Blotting, Western
COS Cells
COUP Transcription Factors
Cell Nucleus / metabolism
DNA / chemistry
DNA-Binding Proteins / metabolism
Gene Expression Regulation
Genes, Reporter
Humans
Interferon Regulatory Factor-1
Luciferases / metabolism
Oligonucleotide Array Sequence Analysis / methods*
Phosphoproteins / metabolism
Plasmids / metabolism
Precipitin Tests
Protein Array Analysis / methods*
Protein Binding
RNA / metabolism
RNA, Small Interfering / metabolism
Receptors, Glucocorticoid / metabolism*
Receptors, Steroid / metabolism
Reverse Transcriptase Polymerase Chain Reaction
TCF Transcription Factors
Transcription Factor 7-Like 1 Protein
Transcription Factors / metabolism*
Transfection

Substances

Actins
COUP Transcription Factors
DNA-Binding Proteins
IRF1 protein, human
Interferon Regulatory Factor-1
Phosphoproteins
RNA, Small Interfering
Receptors, Glucocorticoid
Receptors, Steroid
TCF Transcription Factors
TCF7L1 protein, human
Transcription Factor 7-Like 1 Protein
Transcription Factors
RNA
DNA
Luciferases