Poster abstracts
Poster number 62 submitted by Danushika Herath
CHARACTERIZATION of Transcription Product Outcomes in T-box Riboswitch Modulation
Danushika Herath (Chemistry and Biochemistry, Ohio University), Jennifer V. Hines (Chemistry and Biochemistry, Ohio University)
Abstract:
Developing new antibacterial compounds and finding novel targets is crucial as a solution for antibacterial drug resistance. Non-coding RNA such as T-box riboswitches can be used as efficient drug targets against antibacterial resistance because they regulate amino-acid-related genes essential for bacterial survival. The main objective of this project is to characterize and quantify transcription product outcomes for modulating the T-box riboswitch function. It would provide insights into the RNA structure-function relationships in finding novel drugs against antibacterial resistance. Grundy et al., 2002, discovered that tRNA plays a key role in T-box riboswitch antitermination and this tRNA-dependent antitermination can be studied in vitro in bacterial transcription systems such as Bacillus subtilis by performing in vitro transcription antitermination (IVT) assays. We designed four variants with modified glyQS T-box antiterminator sequences, by mutating the 100% conserved 10th nucleotide position of the 5’- bulge in the glyQS T-box antiterminator of Bacillus subtilis, which is found to be involved in the stacking interactions with the corresponding tRNA. Changes in the transcription antitermination as a result of the specific modifications were assessed through the transcriptional read-through by performing IVT assays. Through these approaches, the role of the specific structural elements in glyQS T-box riboswitch regulation can be elucidated.
Keywords: T-box, Antiterminator, Antibacterial