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Updated: Jun 8, 2026
Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae
Published on: December 29, 2017
A histone deacetylase-dependent screen in yeast
Sujith V W Weerasinghe1, Magdalene Wambua, Mary Kay H Pflum
1Department of Chemistry, 5101 Cass Avenue, Wayne State University, Detroit, MI 48202, USA.
Abstract:
Histone deacetylase (HDAC) proteins are promising targets for cancer treatment, as shown by the recent FDA approval of the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA, Vorinostat) for the treatment of cutaneous T-cell lymphoma. To identify additional potent inhibitors and characterize HDAC mutant proteins, there is interest to develop an inexpensive screening method dependent on native substrates. Here, we report the first yeast-based gene reporter screen dependent on the yeast Rpd3, which is a homolog of human class I HDAC proteins. The screen was sensitive to an inactive Rpd3 mutant and various inhibitors in qualitative, agar-based and quantitative, solution-phase formats. Interestingly, inclusion of the lytic enzyme zymolyase enhanced the inhibitor sensitivity of the screen. The gene reporter screen provides a tool to screen Rpd3 mutants and inhibitors of class I HDAC proteins.
Insights
Researchers developed a novel yeast-based gene reporter screen to identify inhibitors and study mutants of histone deacetylase (HDAC) proteins, specifically targeting class I HDACs like yeast Rpd3. This cost-effective method shows promise for advancing cancer therapeutics research.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Histone deacetylase (HDAC) proteins are crucial in epigenetic regulation and are validated targets for cancer therapy, exemplified by the FDA-approved drug suberoylanilide hydroxamic acid (SAHA).
- Developing efficient screening methods is essential for identifying novel HDAC inhibitors and characterizing HDAC mutant proteins for therapeutic development.
- Yeast Rpd3 is a homolog of human class I HDACs, making it a suitable model for studying HDAC function and inhibition.
Purpose of the Study:
- To establish the first yeast-based gene reporter screen utilizing the yeast Rpd3 protein.
- To develop an inexpensive and sensitive method for screening HDAC inhibitors and characterizing HDAC mutants.
- To assess the utility of the screen in both qualitative and quantitative formats.
Main Methods:
- A yeast-based gene reporter system was engineered to be dependent on the activity of the yeast Rpd3 protein.
- The screen was tested using an inactive Rpd3 mutant and various known HDAC inhibitors.
- Both agar-based (qualitative) and solution-phase (quantitative) screening formats were evaluated.
- The effect of the lytic enzyme zymolyase on inhibitor sensitivity was investigated.
Main Results:
- The developed yeast gene reporter screen demonstrated sensitivity to an inactive Rpd3 mutant, indicating its ability to detect changes in Rpd3 activity.
- The screen successfully identified various HDAC inhibitors in both qualitative and quantitative assays.
- Inclusion of zymolyase significantly enhanced the sensitivity of the screen to HDAC inhibitors.
- The screen provides a robust tool for studying Rpd3 mutants and inhibitors targeting class I HDAC proteins.
Conclusions:
- A novel and cost-effective yeast-based gene reporter screen for class I HDACs, specifically Rpd3, has been successfully developed.
- This screening platform is effective for identifying HDAC inhibitors and characterizing HDAC mutants.
- The enhancement of inhibitor sensitivity by zymolyase offers a valuable optimization for the screening process.
- This tool holds significant potential for advancing the discovery of new cancer therapeutics targeting HDAC proteins.
