Perform a Yeast - three - hybrid screen to identify proteins that bind the SSA4 3' UTR

Publication Date

2026

Presentation Length

Poster/Gallery presentation

College

College of Sciences & Mathematics

Department

Biology, Department of

Student Level

Undergraduate

Faculty Mentor

Rebecca Adams, PhD

Presentation Type

Poster

Summary

During stress conditions like heat shock or ethanol exposure, most mRNA is kept in the nucleus, but some transcripts that help the cell recover are still exported. One of these is SSA4, which codes for a protein that helps refold damaged proteins. It’s still not clear how SSA4 is selectively exported while other mRNAs are not.

We wanted to see if a specific RNA-binding protein interacts with the SSA4 3′ UTR to help it leave the nucleus. We hypothesize that a stress-regulated adaptor protein binds to the SSA4 3′ UTR and allows its selective export.

To test this, we used a yeast three-hybrid (Y3H) screen with the SSA4 3′UTR-MS2 construct and a GAD library to look for protein–RNA interactions under different conditions.

Our results were not usable due to contamination on the plates, which showed abnormal pink coloration and did not smell like yeast. Even though we couldn’t identify any true interactions, this experiment helped set up the process for future trials. Going forward, we plan to repeat the screen using better sterile technique and test both the short and full-length SSA4 3′ UTR to identify potential binding proteins.

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