Strategies to Avoid Artifacts in Mass Spectrometry-Based Epitranscriptome Analyses.

Autor: Kaiser S; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland.; Institute of Pharmaceutical Chemistry, Goethe-University Frankfurt, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany., Byrne SR; Department of Biological Engineering and Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Ammann G; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Asadi Atoi P; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Borland K; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Brecheisen R; Ella Biotech GmbH, 82152, Munich, Germany., DeMott MS; Department of Biological Engineering and Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA., Gehrke T; Ella Biotech GmbH, 82152, Munich, Germany., Hagelskamp F; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Heiss M; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Yoluç Y; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland., Liu L; College of Life Sciences, Qufu Normal University, Qufu, Shandong, 273165, China., Zhang Q; College of Life Sciences, Qufu Normal University, Qufu, Shandong, 273165, China., Dedon PC; Department of Biological Engineering and Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.; Antimicrobial Resistance Interdisciplinary Research Group, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, 138602, Singapore, Singapore., Cao B; College of Life Sciences, Qufu Normal University, Qufu, Shandong, 273165, China., Kellner S; Ludwig-Maximilians Universität München, Butenandtstr. 5-13, 81137, München, Deutschland.; Institute of Pharmaceutical Chemistry, Goethe-University Frankfurt, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2021 Oct 25; Vol. 60 (44), pp. 23885-23893. Date of Electronic Publication: 2021 Sep 29.
DOI: 10.1002/anie.202106215
Abstrakt: In this report, we perform structure validation of recently reported RNA phosphorothioate (PT) modifications, a new set of epitranscriptome marks found in bacteria and eukaryotes including humans. By comparing synthetic PT-containing diribonucleotides with native species in RNA hydrolysates by high-resolution mass spectrometry (MS), metabolic stable isotope labeling, and PT-specific iodine-desulfurization, we disprove the existence of PTs in RNA from E. coli, S. cerevisiae, human cell lines, and mouse brain. Furthermore, we discuss how an MS artifact led to the initial misidentification of 2'-O-methylated diribonucleotides as RNA phosphorothioates. To aid structure validation of new nucleic acid modifications, we present a detailed guideline for MS analysis of RNA hydrolysates, emphasizing how the chosen RNA hydrolysis protocol can be a decisive factor in discovering and quantifying RNA modifications in biological samples.
(© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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