Identifying UTR-Dependent Mechanisms of Selective SSA4 mRNA Export During Heat Stress

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

Identifying UTR-Dependent Mechanisms of Selective SSA4 mRNA Export During Heat Stress

Kyley D. McClung, Rebecca L. Adams PhD

In eukaryotic cells, mRNA export from the nucleus to the cytoplasm is essential for protein synthesis and can be regulated to control gene expression in response to environmental changes. During heat stress at 42°C, most mRNAs are retained in the nucleus while heat shock mRNAs like SSA4 are selectively exported, through the mechanism behind this selective export remains unclear. SSA4 encodes a HSP70 chaperone critical for thermotolerance by refolding damaged proteins during stress.

This study aimed to determine which region of the SSA4 transcript (5’ UTR, ORF, or 3’ UTR) enables selective nuclear export during heat stress. We hypothesized that the 5’ UTR and/or 3’ UTR contain regulatory sequences that facilitate selective export under heat stress conditions. To test this, we used a GFP reporter system fused to SSA4 UTRs and attempted integration at the endogenous SSA4 locus via CRISPR/Cas9, which would allow us to measure GFP expression at normal (25°C) versus heat stress (42°C) temperatures.

I successfully completed plasmid construction, PCR amplification, restriction digest, and yeast transformation steps, but colony PCR screening of 32 transformed colonies did not detect successful integration of the construct. Once these technical issues are resolved, later students can repeat the experiment with better conditions and use the resulting strains to directly compare GFP expression at normal and heat shock temperatures.

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