Institut Pasteur, Department of Structural Biology and Chemistry, Laboratory for Bioorganic Chemistry of Nucleic Acids, CNRS UMR3523, 28, rue du Docteur Roux, 75724 Paris Cedex 15, France.
Université Paris Descartes, Sorbonne Paris Cité, 12 rue de l'École de Médecine, 75006 Paris, France.
ACS Chem Biol. 2020 Nov 20;15(11):2872-2884. doi: 10.1021/acschembio.0c00396. Epub 2020 Oct 22.
The expansion of the genetic alphabet with additional, unnatural base pairs (UBPs) is an important and long-standing goal in synthetic biology. Nucleotides acting as ligands for the coordination of metal cations have advanced as promising candidates for such an expansion of the genetic alphabet. However, the inclusion of artificial metal base pairs in nucleic acids mainly relies on solid-phase synthesis approaches, and very little is known about polymerase-mediated synthesis. Herein, we report the selective and high yielding enzymatic construction of a silver-mediated base pair (dIm-Ag-dPur) as well as a two-step protocol for the synthesis of DNA duplexes containing such an artificial metal base pair. Guided by DFT calculations, we also shed light into the mechanism of formation of this artificial base pair as well as into the structural and energetic preferences. The enzymatic synthesis of the dIm-Ag-dPur artificial metal base pair provides valuable insights for the design of future, more potent systems aiming at expanding the genetic alphabet.
将额外的、非天然碱基对 (UBP) 扩展到遗传密码子是合成生物学中的一个重要且长期存在的目标。作为配位金属阳离子的配体的核苷酸已成为这种遗传密码子扩展的有希望的候选物。然而，人工金属碱基对在核酸中的包含主要依赖于固相合成方法，而对聚合酶介导的合成知之甚少。在此，我们报告了银介导碱基对 (dIm-Ag-dPur) 的选择性和高产酶促构建，以及一种两步法合成含有这种人工金属碱基对的 DNA 双链体的方法。通过 DFT 计算的指导，我们还深入了解了这种人工碱基对的形成机制以及结构和能量偏好。dIm-Ag-dPur 人工金属碱基对的酶促合成为设计旨在扩展遗传密码子的未来更有效系统提供了有价值的见解。
