模板化二氟甲苯核苷酸残基在 RB69 DNA 聚合酶三元复合物中的氢键能力。
Hydrogen-bonding capability of a templating difluorotoluene nucleotide residue in an RB69 DNA polymerase ternary complex.
机构信息
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520, USA.
出版信息
J Am Chem Soc. 2011 Jul 6;133(26):10003-5. doi: 10.1021/ja2021735. Epub 2011 Jun 15.
Abstract
Results obtained using 2,4-difluorotoluene nucleobase (dF) as a nonpolar thymine isostere by Kool and colleagues challenged the Watson-Crick dogma that hydrogen bonds between complementary bases are an absolute requirement for accurate DNA replication. Here, we report crystal structure of an RB69 DNA polymerase L561A/S565G/Y567A triple mutant ternary complex with a templating dF opposite dTTP at 1.8 Å-resolution. In this structure, direct hydrogen bonds were observed between: (i) dF and the incoming dTTP, (ii) dF and residue G568 of the polymerase, and (iii) dF and ordered water molecules surrounding the nascent base pair. Therefore, this structure provides evidence that a templating dF can form novel hydrogen bonds with the incoming dTTP and with the enzyme that differ from those formed with a templating dT.
摘要
Kool 及其同事使用 2,4-二氟甲苯碱基（dF）作为非极性胸腺嘧啶类似物获得的结果，对沃森-克里克碱基互补配对的绝对必要性这一 DNA 复制的经典法则提出了挑战。在此，我们报道了 RB69 DNA 聚合酶 L561A/S565G/Y567A 三突变体与模板 dF 与 dTTP 形成的三元复合物的晶体结构，分辨率为 1.8 Å。在该结构中，观察到直接氢键的形成：（i）dF 与进入的 dTTP，（ii）dF 与聚合酶的残基 G568，以及（iii）dF 与围绕新生碱基对的有序水分子。因此，该结构提供了证据表明，模板 dF 可以与进入的 dTTP 以及与酶形成不同于与模板 dT 形成的氢键。
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