title: Physical manipulation of the Escherichia coli chromosome reveals its soft nature.
abstract: Replicating bacterial chromosomes continuously demix from each other and segregate within a compact volume inside the cell called the nucleoid. Although many proteins involved in this process have been identified, the nature of the global forces that shape and segregate the chromosomes has remained unclear because of limited knowledge of the micromechanical properties of the chromosome. In this work, we demonstrate experimentally the fundamentally soft nature of the bacterial chromosome and the entropic forces that can compact it in a crowded intracellular environment. We developed a unique "micropiston" and measured the force-compression behavior of single Escherichia coli chromosomes in confinement. Our data show that forces on the order of 100 pN and free energies on the order of 10(5) k(B)T are sufficient to compress the chromosome to its in vivo size. For comparison, the pressure required to hold the chromosome at this size is a thousand-fold smaller than the surrounding turgor pressure inside the cell. Furthermore, by manipulation of molecular crowding conditions (entropic forces), we were able to observe in real time fast (approximately 10 s), abrupt, reversible, and repeatable compaction-decompaction cycles of individual chromosomes in confinement. In contrast, we observed much slower dissociation kinetics of a histone-like protein HU from the whole chromosome during its in vivo to in vitro transition. These results for the first time provide quantitative, experimental support for a physical model in which the bacterial chromosome behaves as a loaded entropic spring in vivo.
authors.affiliation: Department of Physics and Section of Molecular Biology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.
authors.forename: James
authors.initials: J
authors.lastname: Pelletier
authors.forename: Ken
authors.initials: K
authors.lastname: Halvorsen
authors.forename: Bae-Yeun
authors.initials: BY
authors.lastname: Ha
authors.forename: Raffaella
authors.initials: R
authors.lastname: Paparcone
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Sandler
authors.forename: Conrad L
authors.initials: CL
authors.lastname: Woldringh
authors.forename: Wesley P
authors.initials: WP
authors.lastname: Wong
authors.forename: Suckjoon
authors.initials: S
authors.lastname: Jun
mesh_terms: D001703:Biophysics; D020090:Chromosome Segregation / Q000502:physiology*; D002876:Chromosomes, Bacterial / Q000737:chemistry / Q000502:physiology*; D019277:Entropy; D004926:Escherichia coli / Q000235:genetics*; D006244:Hardness / Q000502:physiology*; D006245:Hardness Tests / Q000295:instrumentation* / Q000379:methods; D008954:Models, Biological; D011312:Pressure; D013997:Time Factors