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  • In vitro studies of DNA condensation by bridging protein in a crowding environment.

In vitro studies of DNA condensation by bridging protein in a crowding environment.

International journal of biological macromolecules (2017-05-26)
Sravani K Ramisetty, Petter Langlete, Rahmi Lale, Rita S Dias
ABSTRACT

The macromolecules of the bacterial cell occupy 20-40% of the total cytosol volume, and crowded environments have long been known to compact and stabilize DNA. Nevertheless, investigations on DNA-protein binding are generally performed in the absence of crowding, which may yield an incomplete understanding of how nucleoid-assembling proteins work. A family of such proteins, abundant in Gram-negative bacteria, is the histone-like nucleoid structuring proteins (H-NS). Herein, the synergistic role of macromolecular crowding (mimicked using polyethylene glycol, PEG) and H-NS was investigated using fluorescence correlation spectroscopy (FCS) and enzyme protection assays. We show that crowding enhances the binding of H-NS to the AT-rich tracks of the DNA, where it preferentially binds to, protecting these tracks towards enzyme digestion, inducing some DNA condensation, and inhibiting the biological function of DNA. We further suggest that the looping of DNA chains, induced by H-NS, contributes to the synergistic effect of DNA-binding protein and crowding agents, on DNA condensation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-DNA Antibody, double stranded, clone BV16-13, culture supernatant, clone BV16-13, Chemicon®