Poster #054, City of Hope
Characterizing the Physical Interaction Between EPRS1 and miR-146a in a Yeast Three-Hybrid System
Mentors: Thomas Sokolich, PhD, PI: Mark Boldin, MD, PhD
Although inflammation is critical for host defense, its timely resolution is necessary to avoid chronic inflammatory disease. miR-146a serves as a powerful brake on inflammation by post-transcriptionally suppressing the NF-κB pathway. Accordingly, miR-146a-deficient mice develop severe hematological defects, including bone marrow failure, cancer, and multi-organ autoimmunity. miR-146a dysregulation is found in cognate human disorders such as aplastic anemia and autoimmune disease. While most miRNAs rely on canonical biogenesis machinery, our in vitro experiments indicate miR-146a requires additional regulators to fully mature. We identified EPRS1 as a candidate regulator through a miR-146a protein-interactome screen. To confirm this interaction and define the cis-elements involved, I will use pre-miR-146a as bait for EPRS1 in a yeast three-hybrid (Y3H) assay. Consistent with previous results, I hypothesize pre-miR-146a and EPRS1 will bind in yeast. I will also construct a miR-146a dual reporter plasmid to identify additional regulators through genetic screening.