Modified nucleoside triphosphates exist in mammals

Chemical Science
Han-Peng JiangYu-Qi Feng

Abstract

DNA and RNA contain diverse chemical modifications that exert important influences in a variety of cellular processes. In addition to enzyme-mediated modifications of DNA and RNA, previous in vitro studies showed that pre-modified nucleoside triphosphates (NTPs) can be incorporated into DNA and RNA during replication and transcription. Herein, we established a chemical labeling method in combination with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis for the determination of endogenous NTPs in the mammalian cells and tissues. We synthesized 8-(diazomethyl)quinoline (8-DMQ) that could efficiently react with the phosphate group under mild condition to label NTPs. The developed method allowed sensitive detection of NTPs, with the detection limits improved by 56-137 folds. The results showed that 12 types of endogenous modified NTPs were distinctly determined in the mammalian cells and tissues. In addition, the majority of these modified NTPs exhibited significantly decreased contents in human hepatocellular carcinoma (HCC) tissues compared to tumor-adjacent normal tissues. Taken together, our study revealed the widespread existence of various modified NTPs in eukaryotes.

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Citations

Aug 9, 2019·Wiley Interdisciplinary Reviews. RNA·Nils MuthmannAndrea Rentmeister
Sep 24, 2019·Cell Cycle·Boyang Liu, Jörg Großhans
Oct 22, 2020·Biomedical Chromatography : BMC·Victor DavidToma Galaon
Feb 10, 2021·Critical Reviews in Analytical Chemistry·Vladimir G Zaikin, Roman S Borisov
Dec 10, 2020·Science China. Chemistry·Yongxi ZhaoChunhai Fan
Jan 23, 2019·Accounts of Chemical Research·Yafen WangXiang Zhou
Aug 23, 2019·Chemical Research in Toxicology·Bei ChenYu-Qi Feng
Sep 14, 2018·Analytical Chemistry·Bei ChenYu-Qi Feng
Nov 7, 2019·Chemical Research in Toxicology·Bi-Feng Yuan
Mar 29, 2019·Chemical Research in Toxicology·Jun XiongYu-Qi Feng
Jun 13, 2019·ACS Chemical Biology·Cheng-Jie MaYu-Qi Feng
Aug 31, 2021·RSC Chemical Biology·Yi DaiYu-Qi Feng

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Methods Mentioned

BETA
electrophoresis
protein assay
nuclear magnetic resonance
NMR

Software Mentioned

Xcalibur
SPSS

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