speaker Selene Swanson's professional career started in the late 80’s at the Microchemical Facility, University of Minnesota as a junior scientist primarily responsible for protein sequencing, peptide synthesis, DNA sequencing/synthesis and amino acid analysis. After a journey through oncology, hematology, gene therapy and nephrology, she landed in the field of proteomics in the early 2000’s at the Proteomics Center of the Stowers Institute for Medical Research. In 2016, after learning the ropes in MudPIT, she was asked to explore the small molecules arena, specifically modified ribonucleosides, given the availability of state-of-the-art mass spectrometers in the center. With the limited open resources available for analyzing PRM data from small molecules, Skyline has proven to be an invaluable resource to meet our needs.

Absolute quantitative analysis of modified ribonucleosides in tRNA and mRNA using Skyline

Epitranscriptomics is a link between epigenomics and proteomics. Understanding the role of RNA modifications is important in elucidating many fundamental cellular processes such as RNA structural stability, splicing, and translation efficiency. Using a Lumos™ Tribrid™ Mass Spectrometer and Skyline, modified ribonucleosides (RNs) in E. coli tRNA and mammalian mRNA were identified and quantified based on calibration curves generated from known standard RNs. The detection levels of the four 2-O’-methylated RNs ranged from undetectable to 0.7ng and 1.8ng out of 500ng each for mammalian mRNA and tRNA, respectively. Read More
For m7G, an evolutionarily conserved modification of eukaryotic mRNA, 0.46ng was detected from 500ng of hydrolyzed mammalian mRNA. 5’ m7G plays an important role in the life cycle of mRNA such as pre-mRNA processing, nuclear export, and cap-dependent protein synthesis. The ability to identify and quantify these ribonucleoside modifications fosters a mean to compare their abundance level in various cellular and disease states