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Updated: Aug 30, 2025
Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
Published on: November 15, 2013
Simulating androgen receptor selection in designer yeast
Haoran Zhang1,2, Lu Zhang1,2, Yipeng Xu3
1Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, PR China.
Abstract:
Androgen receptor (AR) mutation is closely associated with prostate cancer (PCa) and is one of the mechanisms of resistance to PCa therapies such as AR antagonists. Although sequencing technologies like next-generation sequencing (NGS) contributes to the high-throughput and precise detection of AR mutations carried by PCa patients, the lack of interpretations of these clinical genetic variants has still been a roadblock for PCa-targeted precision medicine. Here, we established a designer yeast reporter assay to simulate natural androgen receptor (AR) selection using AR antagonists. Yeast HIS3 gene transactivation was associated with the ligand-induced recruitment of steroid receptor coactivator-1 (SRC-1) by AR mutants, where yeast growth in histidine-free medium was determined as the outcome. This assay is applicable to determine a wide range of clinical AR mutants including those with loss of function relating to androgen insensitivity syndrome (AIS), and those associated with PCa conferring resistance to AR antagonists such as enzalutamide (ENZ), bicalutamide (BIC), and cyproterone acetate (CPA). One clinical AR mutant previously reported to confer ENZ-resistance, F877L, was found to confer partial resistance to CPA as well using designer yeast. Our simple and efficient assay can enable precise one-pot screening of AR mutants, providing a reference for tailored medicine.
Insights
A new yeast assay accurately screens androgen receptor (AR) mutations in prostate cancer (PCa). This tool aids in understanding treatment resistance and developing personalized medicine for PCa patients.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Androgen receptor (AR) mutations drive prostate cancer (PCa) and therapy resistance.
- Next-generation sequencing (NGS) detects AR mutations, but clinical interpretation remains challenging for precision medicine.
- Understanding AR variant function is crucial for effective PCa treatment strategies.
Purpose of the Study:
- To develop a novel yeast reporter assay for functional screening of AR mutants.
- To simulate AR selection under AR antagonist pressure.
- To provide a tool for interpreting clinical AR variants and guiding precision medicine in PCa.
Main Methods:
- A designer yeast reporter system was established to assess AR mutant activity.
- Yeast growth in histidine-free medium served as a readout for AR transactivation.
- The assay utilized AR antagonists like enzalutamide (ENZ), bicalutamide (BIC), and cyproterone acetate (CPA).
Main Results:
- The yeast assay successfully identified AR mutants conferring resistance to AR antagonists.
- The F877L AR mutant, known to resist ENZ, also showed partial resistance to CPA.
- The assay demonstrated applicability to various AR mutants, including those causing androgen insensitivity syndrome (AIS).
Conclusions:
- A simple and efficient yeast assay enables precise, one-pot screening of AR mutants.
- This assay provides valuable functional insights into AR variants relevant to PCa and treatment resistance.
- The findings offer a reference for developing tailored therapies for prostate cancer patients.
