Poster abstracts
Poster number 64 submitted by Laura Hertz
High-throughput molecular evolution function exploration of the fluoride riboswitch
Laura Marie Hertz (Department of Chemical and Biological Engineering, Northwestern University), Elena Rivas (Department of Molecular and Cellular Biology, Harvard University), Julius B Lucks (Department of Chemical and Biological Engineering, Northwestern University)
Abstract:
RNA structure remains elusive to biophysical techniques due to its flexible backbone and exploration of different ensemble states. However, since the 90s comparative genomics has demonstrated how nucleotides covary within genetic sequences to reveal RNA structures. Here, we expand upon these computational techniques to explore the complete evolution of riboswitches, which are typically cis-regulator RNA elements in the 5` UTR of genes in response to ligand binding. Covariation analysis has only revealed half the structure of a riboswitch, the ligand binding half, due to high variability of structure and function of the gene expression half. We address this gap and then bring comparative genomics out of the computer by developing a high-throughput RNA-sequencing assay to test every unique occurrence of the fluoride riboswitch. We compare the accuracy of computational predictions and validate hits of interest. These findings have led to development of new hypothesizes around the evolution of the RNA sequence-structure-function relationship with implications for biotechnological innovation.
Keywords: riboswitch, structure, high-throughput