Relating Androgen Receptor Conformation to Function in Prostate Cancer Cells
Abstract
Androgen Receptor (AR) recruits SRC coactivators in response to stimulation by hormone and other signals, thereby promoting prostate cancer growth and survival. Activated AR adopts two conformations, a head-to-tail dimer where a coactivator binding site (AF-2) in the C-terminal ligand binding domain (LBD) recognizes a short peptide (FQNLF) in the N-terminal domain of a partner AR and an alternate dimer with AF-2 available for contacts with SRC LxxLL motifs. Here, the authors set out to identify AR mutants committed to particular conformations and to create stable cell lines that express these mutants. As part of an unexpected collaboration, they obtained crystals of AR LBDs in complex with FxxLF and LxxLL peptides and evidence that AR interactions with SRC2 require AF-2 contacts with LxxLL motifs. They learned how to obtain AR mutants unable to form the head-to-tail dimer and created vectors suitable for stable AR expression. They created prostate cancer cells that express a tet-repressor protein and are now selecting cells that express AR under tet control and HeLa cells that express human AR or AR mutants. These cells will be used for studies of effects of AR conformation on coactivator recruitment and prostate cancer cell behavior.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2005
- Accession Number
- ADA434610
Entities
People
- Paul Webb
Organizations
- University of California, San Francisco