Enzymatic Formation of an Artificial Base Pair Using a Modified Purine Nucleoside Triphosphate

Marie Flamme (1) , Pascal Röthlisberger (1) , Fabienne Levi-Acobas (1) , Mohit Chawla (2) , Romina Oliva (3) , Luigi Cavallo (2) , Gilles Gasser (4) , Philippe Marlière (5, 6) , Piet Herdewijn (7) , Marcel Hollenstein (1)

1 Chimie bioorganique des acides nucléiques - Bioorganic chemistry of nucleic acids
2 KAUST - King Abdullah University of Science and Technology [Saudi Arabia]
3 PARTHENOPE - Università degli Studi di Napoli “Parthenope” = University of Naples
4 iCLeHS - Institute of Chemistry for Life and Health Sciences
5 Génopole [Evry]
6 ISSB - Institut de biologie systémique et synthétique
7 Rega Institute for Medical Research [Leuven, België]

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 (dImC–AgI–dPurP) 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 dImC–AgI–dPurP artificial metal base pair provides valuable insights for the design of future, more potent systems aiming at expanding the genetic alphabet.

Domaines: Chimie thérapeutique

Marie Flamme, Pascal Röthlisberger, Fabienne Levi-Acobas, Mohit Chawla, Romina Oliva, et al.. Enzymatic Formation of an Artificial Base Pair Using a Modified Purine Nucleoside Triphosphate. Journal of the American Chemical Society, 2020, 15 (11), pp.2872-2884. ⟨10.1021/acschembio.0c00396⟩. ⟨hal-02975957⟩