Domain interactions between coregulator ARA(70) and the androgen receptor (AR).

The coregulator function of AR-associated protein 70 (ARA(70)) was investigated to further characterize its interaction with the AR. Using a yeast two-hybrid assay, androgen-dependent binding of ARA(70) deletion mutants to the AR ligand-binding domain (LBD) was strongest with ARA(70) amino acids 321-441 of the 614 amino acid ARA(70) protein. Mutations ...
adjacent to or within an FxxLF motif in this 120-amino acid region abolished androgen-dependent binding to the AR-LBD both in yeast and in glutathione-S-transferase affinity matrix assays. Yeast one-hybrid assays revealed an intrinsic ARA(70) transcriptional activation domain within amino acids 296-441. In yeast assays the ARA(70) domains for transcriptional activation and for binding to the AR-LBD were inhibited by the C-terminal region of ARA(70). Full-length ARA(70) increased androgen-dependent AR transactivation in transient cotransfection assays using a mouse mammary tumor virus-luciferase reporter in CV1 cells. ARA(70) also increased constitutive transcriptional activity of an AR NH(2)-terminal-DNA binding domain fragment and bound this region in glutathione-S-transferase affinity matrix assays. Binding was independent of the ARA(70) FxxLF motif. The results identify an ARA(70) motif required for androgen-dependent interaction with the AR-LBD and demonstrate that ARA(70) can interact with the NH(2)-terminal and carboxyl-terminal regions of AR.
Mesh Terms:
Amino Acid Motifs, Androgens, Animals, Binding Sites, Cell Line, Gene Library, Humans, Male, Nuclear Receptor Coactivators, Oncogene Proteins, Protein Binding, Protein Structure, Tertiary, Receptors, Androgen, Receptors, Glucocorticoid, Receptors, Progesterone, Sequence Deletion, Trans-Activators, Transcription Factors, Transcriptional Activation, Transfection, Two-Hybrid System Techniques, Yeasts
Mol. Endocrinol.
Date: Feb. 01, 2002
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