The End Restraint Method for Mechanically Perturbing Nucleic Acids in silico

TL;DR

This paper introduces a flexible in silico umbrella-sampling protocol for mechanically perturbing nucleic acids at the single base pair level, aiding the interpretation of experimental data from optical and magnetic tweezers.

Contribution

It presents a novel, straightforward method for twisting and stretching nucleic acids in simulation, compatible with popular packages like Amber and GROMACS.

Findings

Protocol allows detailed structural analysis of nucleic acids under mechanical stress.

Comparison with experimental data is facilitated through fractional end-to-end displacement.

Method is adaptable for various simulation setups and forcefields.

Abstract

Far from being a passive information store, the genome is a mechanically dynamic and diverse system in which torsion and tension fluctuate and combine to determine structure and help regulate gene expression. Much of this mechanical perturbation is due to molecular machines such as topoisomerases which must stretch and twist DNA as part of various functions including DNA repair and replication. While the broad-scale mechanical response of nucleic acids to tension and torsion is well characterized, detail at the single base pair level is beyond the limits of even super-resolution imaging. Here, we present a straightforward, flexible, and extensible umbrella-sampling protocol to twist and stretch nucleic acids in silico using the popular biomolecular simulation package Amber -- though the principles we describe are applicable also to other packages such as GROMACS. We discuss how to set up the simulation system, decide forcefields and solvation models, and equilibrate. We then introduce the torsionally-constrained stretching protocol, and finally we present some analysis techniques we have used to characterize structural motif formation. Rather than define forces or fictional pseudoatoms, we instead define a fixed translation of specified atoms between each umbrella sampling step, which allows comparison with experiment without needing to estimate applied forces by simply using the fractional end-to-end displacement as a comparison metric. We hope that this easy to implement solution will be valuable for interrogating optical and magnetic tweezers data on nucleic acids at base pair resolution.