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Role of the 2-Amino Group of Purines during dNTP Polymerization by Human DNA Polymerase α

机译：嘌呤的2-氨基在人类DNA聚合酶αdNTP聚合中的作用

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摘要

We used a series of dNTP analogues in conjunction with templates containing modified bases to elucidate the role that N2 of a purine plays during dNTP polymerization by human DNA polymerase α. Removing N2 from dGTP had small effects during correct incorporation opposite C but specifically increased misincorporation opposite A. Adding N2 to dATP and related analogues had small and variable effects on the efficiency of polymerization opposite T. However, the presence of N2 greatly enhanced polymerization of these dATP analogues opposite a template C. The ability of N2 to enhance polymerization opposite C likely results from formation of a hydrogen bond between the purine N2 and pyrimidine O2. Even in those cases where formation of a wobble base pair, tautomerization, and/or protonation of the base pair between the incoming dNTP and template base cannot occur (e.g., 2-pyridone·purine (or purine analogue) base pairs), N2 enhanced formation of the base pair. Importantly, N2 had similar effects on dNTP polymerization both when added to the incoming purine dNTP and when added to the template base being replicated. The mechanistic implications of these results regarding how pol α discriminates between right and wrong dNTPs are discussed.
机译：我们将一系列dNTP类似物与包含修饰碱基的模板结合使用，以阐明嘌呤N2在人DNA聚合酶αdNTP聚合过程中所起的作用。从dGTP中除去N2在正确地掺入C的过程中影响很小，但特别是增加了与A相对的误掺入。向dATP和相关类似物中添加N2对与T相反的聚合效率影响很小且可变。但是，N2的存在极大地增强了这些聚合与模板C相对的dATP类似物。N2增强与C相对的聚合能力的原因可能是嘌呤N2和嘧啶O2之间形成氢键。即使在无法发生摆动碱基对，进入的dNTP和模板碱基之间发生碱基互变异构化和/或碱基对质子化的情况下（例如2-吡啶酮·嘌呤（或嘌呤类似物）碱基对），N2也会增强碱基对的形成。重要的是，N2在添加到传入的嘌呤dNTP中和添加到要复制的模板碱基中时，对dNTP聚合都具有类似的作用。讨论了这些结果关于polα如何区分正确和错误dNTP的机制含义。

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