title: Crosslinking reactions of 4-amino-6-oxo-2-vinylpyrimidine with guanine derivatives and structural analysis of the adducts.
abstract: DNA interstrand crosslinks (ICLs) are the primary mechanism for the cytotoxic activity of many clinical anticancer drugs, and numerous strategies for forming ICLs have been developed. One such method is using crosslink-forming oligonucleotides (CFOs). In this study, we designed a 4-amino-6-oxo-2-vinylpyrimidine (AOVP) derivative with an acyclic spacer to react selectively with guanine. The AOVP CFO exhibited selective crosslinking reactivity with guanine and thymine in DNA, and with guanine in RNA. These crosslinking reactions with guanine were accelerated in the presence of CoCl2, NiCl2, ZnCl2 and MnCl2. In addition, we demonstrated that the AOVP CFO was reactive toward 8-oxoguanine opposite AOVP in the duplex DNA. The structural analysis of each guanine and 8-oxoguanine adduct in the duplex DNA was investigated by high-resolution NMR. The results suggested that AOVP reacts at the N2 amine in guanine and at the N1 or N2 amines in 8-oxoguanine in the duplex DNA. This study demonstrated the first direct determination of the adduct structure in duplex DNA without enzyme digestion.
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai-shi, Miyagi 980-8577, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Kusano
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai-shi, Miyagi 980-8577, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Ishiyama
authors.affiliation: Department of Chemistry, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China lams@cuhk.edu.hk.
authors.forename: Sik Lok
authors.initials: SL
authors.lastname: Lam
authors.affiliation: Institute of Advanced Energy, Graduate School of Energy Science, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Mashima
authors.affiliation: Institute of Advanced Energy, Graduate School of Energy Science, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Katahira
authors.affiliation: Department of Chemistry, Graduate School of Science, Hiroshima University,1-3-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan.
authors.forename: Kengo
authors.initials: K
authors.lastname: Miyamoto
authors.affiliation: Department of Chemistry, Graduate School of Science, Hiroshima University,1-3-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan.
authors.forename: Misako
authors.initials: M
authors.lastname: Aida
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai-shi, Miyagi 980-8577, Japan nagatugi@tagen.tohoku.ac.jp.
authors.forename: Fumi
authors.initials: F
authors.lastname: Nagatsugi
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry*; D004247:DNA / Q000737:chemistry; D018736:DNA Adducts / Q000737:chemistry*; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis; D011744:Pyrimidinones / Q000737:chemistry*; D012313:RNA / Q000737:chemistry; D013941:Thymine / Q000737:chemistry; D014753:Vinyl Compounds / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D018736:DNA Adducts; D008670:Metals; D009841:Oligonucleotides; D011744:Pyrimidinones; D014753:Vinyl Compounds; D006147:Guanine; D012313:RNA; D004247:DNA; D013941:Thymine