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Global Profiling of the Oxidative Stress Induced Effects on RNA Modifications by Liquid Chromatography‐Tandem Mass Spectrometry
Author(s) -
Jora Manasses,
Sun Congliang,
Limbach Patrick A.,
Addepalli Balasubrahmanyam
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.787.7
Subject(s) - chemistry , oxidative stress , oxidative phosphorylation , rna , reactive oxygen species , transfer rna , photosensitizer , dna damage , tandem mass spectrometry , nucleoside , liquid chromatography–mass spectrometry , biochemistry , mass spectrometry , dna , biophysics , chromatography , photochemistry , gene , biology
The goal of this work was to profile the oxidative damage exerted by ultraviolet‐A radiation (UVA, 370 nm) or H 2 O 2 on RNA modifications by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Oxidative damage to RNA impacts biological systems due to its association with metabolic, mitochondrial, neurological diseases and cancer.1,2 In the presence of photosensitizer ( e.g. , riboflavin), UVA radiation induces the production of reactive oxygen species (ROS, O 2 • , HOO • , HO • ) in vivo .3 Likewise, H 2 O 2 generates ROS under physiological conditions, and formation of radicals is accelerated via Fenton reactions in the presence of catalysts ( e.g. , Fe 2+ , Cu 2+ ).4 Oxidation pathways of canonical nucleosides are well documented.5 However, the effect of ROS on modified ribonucleosides is not well known, and information about RNA damage is largely extrapolated from studies on DNA. More than 160 modified nucleosides have been reported in various types of RNA of different organisms.6 E. coli with 46 modifications in its tRNA serve as a good model system to investigate the stress‐induced effects. Apart from the known oxidative products arising from canonical nucleosides, we have documented a list of modified nucleosides that are adversely affected by UVA or H 2 O 2 . To understand the oxidative environment generated by UVA exposure, we exposed E. coli total tRNA with or without a photosensitizer for defined period of time. In a similar manner, the effect of H 2 O 2 on RNA modifications was investigated by incubating E. coli total tRNA and H 2 O 2 , with or without the presence of transition metal ion catalysts. The oxidative damage observed on various modifications was measured by subjecting it to LC‐MS/MS following complete hydrolysis of tRNA. We will present data that describes the susceptibility of various functional groups of these modifications and underlying patterns. Further, these studies also document a range of RNA photoproducts and chemical alterations arising due to the oxidative conditions. Support or Funding Information Financial support of this work was provided by the NSF (CHE1507357) and DTRA (HDTRA1‐15‐1‐0033), both to Patrick A. Limbach, and the University of Cincinnati. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .