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The volume occupied by unconstrained bacterial DNA in physiological solutionsexceeds 1000 times the volume of the cell. Still, it is confined to a welldefined region of the cell called the nucleoid, which occupies only a fractionof the cell volume. There is still no general agreement on the mechanismleading to the compaction of the DNA and the formation of the nucleoid.However, advances in in vivo sub-wavelength resolution microscopy techniqueshave recently allowed the observation of the nucleoid at an unprecedented levelof detail. In particular, these observations show that the compaction of thenucleoid is not static but is instead a highly dynamic feature, which dependson several factors, like the richness of the nutrient, the cell cycle stage,temperature, the action of an osmotic shock or antibiotics, etc. After a shortdescription of the electrolyte content of the cytosol and a brief overview ofthe different mechanisms that may lead to the formation of the nucleoid, thispaper reviews some of the most fascinating recent results of in vivosub-wavelength resolution microscopy. It is furthermore argued that theseobservations provide converging indications in favor of a model that describesthe cytosol as an aqueous electrolyte solution containing severalmacromolecular species, where demixing and segregative phase separation occurbetween DNA and RNA (essentially rRNA and mRNA involved in translationcomplexes, but also the large amounts of rRNA synthesized at the rrn operons ofcells growing in rich media). It is also pointed out that crowding may play acrucial role through its synergy with electrostatic forces. By constrainingmacromolecules to remain at short distances from one another and feelelectrostatic interactions in spite of the strong screening exerted byelectrolyte species, crowding favors stronger DNA/RNA demixing and nucleoidcompaction.

In vivo compaction dynamics of bacterial DNA: A fingerprint of DNA/RNA demixing ?