444 Post-translational role of RNA modifications in sRNA chaperone Hfq

OBJECTIVES/GOALS: The goal of this study is to determine the role of the tRNA modifications in the translation of Hfq. Hfq is an RNA chaperone that acts as a co-factor for the action of the largest class of small RNAs in E. coli. RNA modifications have been known to play critical roles in the translational fidelity of many cellular proteins in bacteria. METHODS/STUDY POPULATION: In this study, we used an hfq-lacZ translation fusion to screen several RNA modification mutant genes to uncover additional RNA modifications that may play a role in Hfq translation. We measured hfq-lacZ activity in genetic backgrounds mutated for several additional RNA modification enzymes previously untested for Hfq effects. RESULTS/ANTICIPATED RESULTS: We identified 5 RNA modification genes that were defective for hfq-lacZ fusion activity, and we subsequently performed western blot analysis on the Hfq protein in the absence of these modification mutant genes to determine the effect of these mutants more directly on Hfq protein levels. We identified 2 out of these 5 RNA modification mutants that also affect Hfq protein levels. DISCUSSION/SIGNIFICANCE: Since Hfq is critically important for small RNA function is a wide range of bacteria, it is possible tRNA modifications regulate Hfq expression in other bacteria. These processes, when further investigated, could provide us with the basic information to develop new antibiotics needed to address emerging antibiotic resistance.