Genome-wide DNA bridging by H-NS reshapes the stationary phase nucleoid and transcriptional landscape

Bacterial nucleoid-associated proteins (NAPs) structure the chromosome and regulate gene expression, but how these two functions are related is unclear. H-NS is a well-studied NAP that acts as a global gene silencer capable of bridging and looping DNA in vitro . Here, using high-throughput chromosome conformation capture assays, we show that H-NS mediates genome-wide long-range DNA looping in the stationary-phase nucleoid of Escherichia coli . Chromatin immunoprecipitation assays demonstrate that high levels of H-NS are present at the base of DNA loops. Super-resolution imaging and single-particle tracking show that H-NS binds more tightly in stationary phase and compacts the nucleoid mesh. Transcriptomic analyses indicate H-NS represses gene expression more strongly in the looped nucleoid and enables higher expression of genes outside of H-NS-bound regions. Overall, our study demonstrates that H-NS bridges distal DNA regions along the genome upon nutrient limitation, causing reduced nucleoid accessibility, stronger transcriptional repression, and a shifted transcriptional landscape.