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Mating-based Overexpression Library Screening in Yeast
Published on: July 6, 2018
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Mutant screening for oncogenes of Ewing's sarcoma using yeast
Takao Kitagawa1, Hajime Okita, Byron Baron
1Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi, 755-8505, Japan, takao-k@yamaguchi-u.ac.jp.
Applied Microbiology and Biotechnology
|May 5, 2015
Summary
A novel yeast screening identified key regions in Ewing sarcoma oncogenes essential for their function. Mutations within the ETS domain of EWS/ETS proteins suppressed growth inhibition, highlighting it as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Fusion genes resulting from chromosomal translocations act as oncogenes.
- The precise mechanisms of action for many oncogenes remain unclear.
- Ewing sarcoma is driven by specific fusion oncogenes.
Purpose of the Study:
- To identify essential functional regions of Ewing sarcoma oncogenes.
- To develop a yeast-based screening system for oncogene function.
- To investigate the role of specific domains in oncogenic activity.
Main Methods:
- Utilized a yeast mutant screening system to assess oncogene function.
- Introduced specific oncogenes (EWS/FLI1, EWS/ERG, EWS/E1AF) into yeast.
- Identified single amino acid substitution mutants that suppressed oncogene-induced growth inhibition.
Main Results:
- Discovered 13 single amino acid substitution mutants that alleviated growth inhibition.
- Localized critical mutation sites to the ETS domain of EWS/ETS proteins.
- Observed reduced transcriptional activity of mutated proteins on the DKK2 promoter.
Conclusions:
- The ETS domain of EWS/ETS family proteins is crucial for Ewing sarcoma growth inhibition.
- The identified yeast screening system is effective for functional screening of oncogenes.
- Targeting the ETS domain presents a potential therapeutic strategy for Ewing sarcoma.
