Next-generation sequencing reveals the biological significance of the N[superscript 2],3-ethenoguanine lesion in vivo
Author(s)Chang, Shiou-chi; Fedeles, Bogdan I.; Wu, Jie; Delaney, James C.; Li, Deyu; Zhao, Linlin; Christov, Plamen P.; Yau, Emily; Singh, Vipender; Jost, Marco; Drennan, Catherine L.; Marnett, Lawrence J.; Rizzo, Carmelo J.; Levine, Stuart S.; Guengerich, F. Peter; Essigmann, John M.; ... Show more Show less
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Etheno DNA adducts are a prevalent type of DNA damage caused by vinyl chloride (VC) exposure and oxidative stress. Etheno adducts are mutagenic and may contribute to the initiation of several pathologies; thus, elucidating the pathways by which they induce cellular transformation is critical. Although N[superscript 2],3-ethenoguanine (N[superscript 2],3-εG) is the most abundant etheno adduct, its biological consequences have not been well characterized in cells due to its labile glycosidic bond. Here, a stabilized 2′-fluoro-2′-deoxyribose analog of N[superscript 2],3-εG was used to quantify directly its genotoxicity and mutagenicity. A multiplex method involving next-generation sequencing enabled a large-scale in vivo analysis, in which both N[superscript 2],3-εG and its isomer 1,N[superscript 2]-ethenoguanine (1,N[superscript 2]-εG) were evaluated in various repair and replication backgrounds. We found that N[superscript 2],3-εG potently induces G to A transitions, the same mutation previously observed in VC-associated tumors. By contrast, 1,N[superscript 2]-εG induces various substitutions and frameshifts. We also found that N[superscript 2],3-εG is the only etheno lesion that cannot be repaired by AlkB, which partially explains its persistence. Both εG lesions are strong replication blocks and DinB, a translesion polymerase, facilitates the mutagenic bypass of both lesions. Collectively, our results indicate that N[superscript 2],3-εG is a biologically important lesion and may have a functional role in VC-induced or inflammation-driven carcinogenesis.
DepartmentMassachusetts Institute of Technology. Center for Environmental Health Sciences; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemical Engineering; Massachusetts Institute of Technology. Department of Chemistry
Nucleic Acids Research
Oxford University Press
Chang, Shiou-chi, Bogdan I. Fedeles, Jie Wu, James C. Delaney, Deyu Li, Linlin Zhao, Plamen P. Christov, et al. “Next-Generation Sequencing Reveals the Biological Significance of the N[superscript 2],3-Ethenoguanine Lesion in Vivo.” Nucleic Acids Research (April 2, 2015).
Final published version