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Updated: Sep 27, 2026
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
A yeast-based functional assay for the detection of the mutant androgen receptor in prostate cancer
Jocelyn Céraline1, Eva Erdmann, Philippe Erbs
1Laboratoire de Cancerologie Experimentale et de Radiobiologie, EA 3430-ULP, IRCAD, 1, Place de l'Hopital, BP426, F67091 Strasbourg, France. jocelyn.ceraline@ircad.u-strasbg.fr
Objective:
Mutations in the ligand-binding domain of the human androgen receptor (AR) figure among the ways used by prostate adenocarcinoma (PCa) cells to escape androgen dependence. These mutations may broaden the specificity and/or affinity of the AR to other hormones, resulting in inappropriate receptor activation and thus affecting the PCa response to physiological stimuli and hormonal therapies.
Design:
In order to clarify the impact of these mutations on disease progression and treatment, we have developed a yeast-based functional assay that allows the detection of mutant ARs and the analysis of their transactivation capacities in response to different ligands.
Methods:
AR cDNA was directly cloned into an expression vector in a yeast strain that carries a reporter gene (ADE2) linked to an androgen-dependent promoter. The expression of the ADE2 gene and consequently the yeast cell growth in a selective medium depleted in adenine depends on the specificity of the AR for the ligand added to the medium.
Results:
By analysing the transactivation capacities of different AR molecules in response to a broad range of steroid and non-steroid ligands, we have demonstrated that this assay can discriminate among wild-type AR, T877A, C685Y and L701H mutant ARs and that at least 1% of mutant ARs could be detected when mutant and wild-type ARs were mixed at the cDNA level.
Conclusions:
The data presented here show that this simple AR assay is convenient for the routine detection of mutant ARs in PCa and is also suitable to evaluate the antagonist activities of anti-androgen molecules.
Insights
This study presents a new yeast-based assay to detect mutant androgen receptors (ARs) in prostate cancer (PCa). This method can identify AR mutations that drive cancer growth and assess anti-androgen drug effectiveness.
Area of Science:
- Molecular biology
- Cancer research
- Endocrinology
Background:
- Mutations in the androgen receptor (AR) ligand-binding domain allow prostate adenocarcinoma (PCa) to evade androgen dependence.
- These AR mutations can alter hormone specificity and affinity, leading to aberrant receptor activation and impacting treatment efficacy.
Purpose of the Study:
- To develop and validate a yeast-based functional assay for detecting mutant ARs.
- To analyze the transactivation capacities of mutant ARs in response to various ligands.
- To assess the utility of the assay for routine detection and evaluating anti-androgen activity.
Main Methods:
- Cloning of AR cDNA into a yeast expression vector with an ADE2 reporter gene.
- Utilizing a yeast strain with an androgen-dependent promoter linked to the ADE2 reporter.
- Assessing yeast growth in selective media based on AR-ligand specificity.
Main Results:
- The assay successfully discriminated between wild-type AR and specific mutants (T877A, C685Y, L701H).
- The assay demonstrated sensitivity, detecting at least 1% mutant ARs in mixed cDNA samples.
- Transactivation capacities of different AR variants were analyzed across a wide range of ligands.
Conclusions:
- The developed yeast assay is a simple and convenient tool for routine detection of mutant ARs in PCa.
- The assay is suitable for evaluating the antagonist activities of anti-androgen compounds.
- This method aids in understanding AR mutation impact on PCa progression and treatment response.
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