Ethan Jeong Rose Hills
A High-Throughput Screening of a Transcriptional Activation Domain
Cellular development relies on transcription factors, specifically their Activation Domains (ADs), to regulate gene expression. Because ADs lack a fixed 3D shape, researchers have long struggled to understand their mechanism of action. The Acidic Exposure Model suggests that ADs transition between a “collapsed” inactive state and an “expanded” active state, where the amount of time spent in the expanded state determines their activity. Currently, measuring these movements is limited to single-molecule techniques. This project aims to develop a high-throughput method using split green fluorescent protein (GFP) to quantify these dynamics. By attaching split GFP fragments to the ADs, we can correlate fluorescence with protein collapse: as the AD folds, the GFP pieces reconstitute into a fluorescent molecule. After validating this method in S. cerevisiae, we will apply it to a vast library of AD variants to map how specific chemical properties, such as acidity and hydrophobicity, control these opening and closing kinetics. This research will provide a scalable, quantitative framework to understand the relationship between protein structure and gene regulation.
Message To Sponsor
I am Ethan, a rising junior studying Chemical Biology. I would like to thank you for this opportunity allowing me to pursue biophysics research in the Bustamante group this summer. I am eager to contribute to the field of transcription factors, which has broad implications in molecular biology and medicine.