Poster abstracts
Poster number 3 submitted by Rachel Braun
Fine-tuning a novel fluorescence assay for the high-throughput screening of RNase P
Rachel M. Braun (Department of Chemistry & Biochemistry and Center for RNA Biology, The Ohio State University), Edric K. Choi (Department of Chemistry & Biochemistry and Center for RNA Biology, The Ohio State University), Sravya Kovvali (Department of Chemistry & Biochemistry and Center for RNA Biology, The Ohio State University), Venkat Gopalan (Department of Chemistry & Biochemistry and Center for RNA Biology, The Ohio State University)
Abstract:
Ribonuclease P (RNase P) is an essential enzyme that catalyzes the 5' maturation of transfer RNAs (tRNAs). The ancient form of RNase P is a ribonucleoprotein and belongs to a select group of biological catalysts that use a catalytic RNA to accomplish function in vivo1. The striking diversity in the protein subunit composition of the RNP forms in the three domains of life has inspired two types of studies: understanding why a catalytic RNA relies on a variable number of protein cofactors in different life forms, and evaluating if these differences can be exploited to render RNase P a viable drug target. The latter goal, however, necessitates high-throughput, economical assays to replace the current radioisotope-based, low-throughput version. In this regard, we have recently developed a sensitive turn-on assay wherein there is a strong fluorescence gain upon RNase P cleavage-mediated release of a fluorescence quencher-coupled oligonucleotide, which is initially annealed to the 5' leader of a 3'-fluor-labeled precursor-tRNA substrate. Results that provide proof-of-principle and establish the bona fides of this novel assay will be presented.
References:
[1]Altman, S (2011) Phil. Trans. R. Soc. B. 366, 2936-2941.
Keywords: RNase P, HTS of an appealing drug target