Characterizing interaction of multiple nanocavity confined plasmids in presence of large DNA model nucleoid

Bacteria have numerous large dsDNA molecules that freely interact within the cell, including multiple plasmids, primary and secondary chromosomes. The cell membrane maintains a micron-scale confinement, ensuring that the dsDNA species are proximal at all times and interact strongly in a manner influenced by the cell morphology (e.g. whether cell geometry is spherical or anisotropic). These interactions lead to non-uniform spatial organization and complex dynamics, including segregation of plasmid DNA to polar and membrane proximal regions. However, exactly how this organization arises, how it depends on cell morphology and number of interacting dsDNA species are under debate. Here, using an in vitro nanofluidic model, featuring a cavity that can be opened and closed in situ, we address how plasmid copy number and confinement geometry alter plasmid spatial distribution and dynamics. We find that increasing the plasmid number alters the plasmid spatial distribution and shortens the plasmid polar dwell time; sharper cavity end curvature leads to longer plasmid dwell times.

Automated summary

Bacteria have various large dsDNA molecules that interact within the cell and are affected by cell morphology, leading to non-uniform spatial organization and dynamics. This study investigates how plasmid copy number and confinement geometry affect plasmid distribution and dynamics using an in vitro nanofluidic model. The results show that increased plasmid number changes distribution and reduces polar dwell time, while sharper cavity end curvature leads to longer dwell time.