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Cooperative effects on the compaction of DNA fragments by the nucleoid protein H-NS and the crowding agent PEG probed by Magnetic Tweezers

TitleCooperative effects on the compaction of DNA fragments by the nucleoid protein H-NS and the crowding agent PEG probed by Magnetic Tweezers
Publication TypeJournal Article
Year of Publication2020
AuthorsCristofalo, M, Marrano, CA, Salerno, D, Corti, R, Cassina, V, Sclavi, B, Gherardi, M, Mammola, A, M. Lagomarsino, C, Mantegazza, F
JournalBiochimica et Biophysica Acta (BBA) - General Subjects
Volume1864
Issue12
Pagination129725
ISSN0304-4165
KeywordsForce spectroscopy, H-NS, Magnetic Tweezers, Nucleoid-associated proteins, Single molecule
Abstract

Background DNA bridging promoted by the H-NS protein, combined with the compaction induced by cellular crowding, plays a major role in the structuring of the E. coli genome. However, only few studies consider the effects of the physical interplay of these two factors in a controlled environment. Methods We apply a single molecule technique (Magnetic Tweezers) to study the nanomechanics of compaction and folding kinetics of a 6 kb DNA fragment, induced by H-NS bridging and/or PEG crowding. Results In the presence of H-NS alone, the DNA shows a step-wise collapse driven by the formation of multiple bridges, and little variations in the H-NS concentration-dependent unfolding force. Conversely, the DNA collapse force observed with PEG was highly dependent on the volume fraction of the crowding agent. The two limit cases were interpreted considering the models of loop formation in a pulled chain and pulling of an equilibrium globule respectively. Conclusions We observed an evident cooperative effect between H-NS activity and the depletion of forces induced by PEG. General Significance Our data suggest a double role for H-NS in enhancing compaction while forming specific loops, which could be crucial in vivo for defining specific mesoscale domains in chromosomal regions in response to environmental changes.

URLhttp://www.sciencedirect.com/science/article/pii/S0304416520302373
DOI10.1016/j.bbagen.2020.129725

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