Poster abstracts

Poster number 175 submitted by Ruoxia Zhao

Detection of Post-Transcriptional RNA Modifications in the Radioresistant Bacterium Deinococcus radiodurans R1

Ruoxia Zhao (Department of Chemistry, Rieveschl Laboratories for Mass Spectrometry, University of Cincinnati), Balasubrahmanyam Addepalli (Department of Chemistry, Rieveschl Laboratories for Mass Spectrometry, University of Cincinnati), Patrick A. Limbach (Department of Chemistry, Rieveschl Laboratories for Mass Spectrometry, University of Cincinnati)

Abstract:
The bacterium Deinococcus radiodurans is one of the most ionizing radiation resistant families of bacteria discovered to date. D. radiodurans has extraordinary resistance to all reactive oxygen species(ROS)-generating agent including desiccation, ionizing radiation, UV radiation, mitomycin C and so on.
In order to understand whether transfer and ribosomal RNAs (tRNAs, rRNAs) are modified differently when D. radiodurans are exposed to ionizing radiation (IR), the modified nucleosides in control D. radiodurans and D. radiodurans with different extent of IR (60Co) have been analyzed by Reverse-Phase High Performance Liquid Chromatography Mass Spectrometry (RP-HPLC-MS/MS). RNA modification mapping is conducted by utilizing a base-specific ribonuclease (RNase T1) to cleave a larger RNA into smaller digestion products (oligonucleotides). The location and identity of nucleotides and modified nucleotides can be determined by using of a DNA-based exclusion list which enhances data dependent RP-HPLC-MS/MS detection method. Determining the modified RNA sequence will increase the understanding of structure-function relationships of RNAs.

References:
Battista, J. R. 1997. Against all odds: the survival strategies of Deinococcus radiodurans. Annu. Rev. Microbiol. 51:203–224.

Battista, J. R., A. M. Earl, and M. J. Park. 1999. Why is Deinococcus radiodurans so resistant to ionizing radiation? Trends Microbiol 7:362-5.
Ross, Robert, et al. "Sequence mapping of transfer RNA chemical modifications by liquid chromatography tandem mass spectrometry." Methods (2016).
Cao, Xiaoyu, and Patrick A. Limbach. "Enhanced detection of post-transcriptional modifications using a mass-exclusion list strategy for RNA modification mapping by LC-MS/MS." Analytical chemistry 87.16 (2015): 8433-8440.

Keywords: Radioresistant, RNA modifications, HPLC-MSMS