Crowding induces entropically-driven changes to DNA dynamics that depend on crowder structure and ionic conditions

TL;DR

This study investigates how macromolecular crowding influences DNA dynamics, revealing that crowder structure and ionic conditions significantly affect DNA conformation and mobility through entropic effects, with implications for biological processes.

Contribution

The paper provides experimental evidence on how different crowder structures and ionic conditions modulate DNA dynamics, highlighting entropic effects in crowded environments.

Findings

Branched crowders compact DNA, linear crowders elongate DNA.

DNA mobility reduction is largely insensitive to crowder structure.

DNA diffusion and size show non-monotonic dependence on salt concentration.

Abstract

Macromolecular crowding plays a principal role in a wide range of biological processes including gene expression, chromosomal compaction, and viral infection. However, the impact that crowding has on the dynamics of nucleic acids remains a topic of debate. To address this problem, we use single-molecule fluorescence microscopy and custom particle-tracking algorithms to investigate the impact of varying macromolecular crowding conditions on the transport and conformational dynamics of large DNA molecules. Specifically, we measure the mean-squared center-of-mass displacements, as well as the conformational size, shape, and fluctuations, of individual 115 kbp DNA molecules diffusing through various in vitro solutions of crowding polymers. We determine the role of crowder structure and concentration, as well as ionic conditions, on the diffusion and configurational dynamics of DNA. We find that branched, compact crowders (10 kDa PEG, 420 kDa Ficoll) drive DNA to compact, whereas linear, flexible crowders (10 kDa, 500 kDa dextran) cause DNA to elongate. Interestingly, the extent to which DNA mobility is reduced by increasing crowder concentrations appears largely insensitive to crowder structure (branched vs linear), despite the highly different configurations DNA assumes in each case. We also characterize the role of ionic conditions on crowding-induced DNA dynamics. We show that both DNA diffusion and conformational size exhibit an emergent non-monotonic dependence on salt concentration that is not seen in the absence of crowders.