Hydrophobic unnatural base pairs show a Watson-Crick pairing in micro-second molecular dynamics simulations

Two unnatural hydrophobic nucleotides, d5SICS (2,6-dimethyl-2H-isoquiniline-1-thione) and dNaM (2-methoxy-3-methylnaphthalene), were previously replicated in vivo by a modified E. coli strand, however, a consistent structure for their pairing in terms of specific and selective directional interactions remains elusive, as data from spectroscopy experiments and simulations are inconsistent. The proposed d5SICS-dNaM pairing has been suggested to be a stacked configuration as suggested by NMR data; simulations have failed to reproduce this configuration and a Watson-Crick like pairing is observed. Previously, we focused on reproducing the d5SICS-dNaM Unnatural Base Pair (UBP) paring using an older (bsc0) AMBER force field, which was not able to correctly reproduce the experimental data. We present our efforts to reproduce the experimental pairing using the current version of the AMBER DNA force fields (OL15 and bsc1), two water models (OPC and TIP3P) and external electrostatic stabilization by Mg²⁺ ions. Opposite to previously reported simulations, a Watson-Crick-like pairing with no hydrogen bonds persists throughout all our results. Despite our efforts to replicate the reported stacked conformation, we cannot confirm its plausibility nor obtain a consistent structure that is independent of the neighboring nucleotides. Communicated by Ramaswamy H. Sarma.