Interpretation of organizational role of proteins on E. coli nucleoid via Hi-C integrated model

Abstract

Several proteins in Escherichia coli work together to maintain the complex organization of its chromosome. However, the individual roles of these so-called nucleoid-associated proteins (NAPs) in chromosome architectures are not well characterized. Here, we quantitatively dissect the organizational roles of Heat Unstable (HU), a ubiquitous protein in E. coli and MatP, an NAP specifically binding to the Ter macrodomain of the chromosome. Toward this end, we employ a polymer physics-based computer model of wild-type chromosome and their HU- and MatP-devoid counterparts by incorporating their respective experimentally derived Hi-C contact matrix, cell dimensions, and replication status of the chromosome commensurate with corresponding growth conditions. Specifically, our model for the HU-devoid chromosome corroborates well with the microscopy observation of compaction of chromosome at short genomic range but diminished long-range interactions, justifying precedent hypothesis of segregation defect upon HU removal. Control simulations point out that the change in cell dimension and chromosome content in the process of HU removal holds the key to the observed differences in chromosome architecture between wild-type and HU-devoid cells. On the other hand, simulation of MatP-devoid chromosome led to locally enhanced contacts between Ter and its flanking macrodomains, consistent with previous recombination assay experiments and MatP's role in insulation of the Ter macrodomain from the rest of the chromosome. However, the simulation indicated no change in matS sites' localization. Rather, a set of designed control simulations showed that insulation of Ter is not caused by bridging of distant matS sites, also lending credence to a recent mobility experiment on various loci of the E. coli chromosome. Together, the investigations highlight the ability of an integrative model of the bacterial genome in elucidating the role of NAPs and in reconciling multiple experimental observations.