Functional Genetic Analysis Reveals Intricate Roles of Conserved X-box Elements in Yeast Transcriptional Regulation

• Main Author: Voll, Sarah
• Other Authors: Kaern, Mads
• Language: en
• Published: Université d'Ottawa / University of Ottawa 2013
• Subjects: functional genetic analysis; transcription factor (TF); cis-regulatory element (CRE); binary interaction; higher order interaction; multiple linear regression; additive; multiplicative; synthetic genetic array (SGA); green fluorescent protein (GFP); ribonucleotide reductase (RNR); yeast; X-box; transcriptional regulation; RNR3
• Online Access: http://hdl.handle.net/10393/30168; http://dx.doi.org/10.20381/ruor-3364

Understanding the functional impact of physical interactions between proteins and DNA on gene expression is important for developing approaches to correct disease-associated gene dysregulation. I conducted a systematic, functional genetic analysis of protein-DNA interactions in the promoter region of the yeast ribonucleotide reductase subunit gene RNR3. I measured the transcriptional impact of systematically perturbing the major transcriptional regulator, Crt1, and three X-box sites on the DNA known to physically bind Crt1. This analysis revealed interactions between two of the three X-boxes in the presence of Crt1, and unexpectedly, a significant functional role of the X-boxes in the absence of Crt1. Further analysis revealed Crt1- independent regulators of RNR3 that were impacted by X-box perturbation. Taken together, these results support the notion that higher-order X-box-mediated interactions are important for RNR3 transcription, and that the X-boxes have unexpected roles in the regulation of RNR3 transcription that extend beyond their interaction with Crt1.