Synthesis and characterization of oligodeoxynucleotides containing formamidopyrimidine lesions and nonhydrolyzable analogues
Synthesis and characterization of oligodeoxynucleotides containing formamidopyrimidine lesions and nonhydrolyzable analogues
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DOI：
10.1021/ja012135q
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发表时间：
2002-04-03
影响因子：
15
通讯作者：
Greenberg, MM
中科院分区：
文献类型：
--
作者：
Haraguchi, K;Delaney, MO;Greenberg, MM
通过固相合成和索马酶促连接法制备了在特定位点含有甲酰胺嘧啶损伤和C-核苷类似物的寡脱氧核苷酸。甲酰胺基嘧啶损伤作为二核苷酸引入，以防止重排为它们的吡喃糖异构体。含有Fady（.）的C-核苷类似物的单一非对映异构体的寡脱氧核苷酸通过使用相应的亚磷酰胺引入dA。甲酰胺基嘧啶病变减少T的十二聚体相对于其未修饰的核苷酸对应物时，相对于核苷酸正确的碱基配对的合作伙伴。然而，含有FcB（.）dG-dA错配对的解链显著高于由dG-dA组成的那些。dA-dX或其C-核苷类似物的熔点低于含有dA-dX的相应复合物。含甲酰胺基嘧啶损伤的寡脱氧核苷酸的碱不稳定性的研究表明，FRENTA（.）使用哌啶（1.0 M，90 ℃，20 min），dA很容易被鉴定为碱不稳定损伤，但使用氟哌啶（.）dG不太容易以这种方式识别。
Oligodeoxynucleotides containing formamidopyrimidine lesions and C-nucleoside analogues at defined sites were prepared by solid-phase synthesis and in soma oases enzymatic ligation. Formamidopyrimidine lesions were introduced as dinucleotides to prevent rearrangement to their pyranose isomers. Oligodeoxynucleotides containing single diastereomers of C-nucleoside analogues of Fady(.)dA were introduced by using the respective phosphoramidites. The formamidopyrimidine lesions reduce the T of dodecamers relative to their unmodified nucleotide counterparts when opposite the nucleotide proper base-pairing partner. However, duplexes containing Fapy(.)dG-dA mispairs melt significantly higher than those comprised of dG-dA. All duplexes containing Fapy(.)dA-dX or its C-nucleoside analogue melt lower than the respective complexes containing dA-dX. Studies of the alkaline lability of oligodeoxynucleotides containing formamidopyrimidine lesions indicate that Fapy(.)dA is readily identified as an alkali-labile lesion with use of piperidine (1.0 M, 90 degreesC, 20 min), but Fapy(.)dG is less easily identified in this manner.