Internal dynamics of supercoiled DNA molecules.
The intramolecular diffusive motion within supercoiled DNA molecules is of central importance for a wide array of gene regulation processes. It has recently been shown, using fluorescence correlation spectroscopy, that plasmid DNA exhibits unexpected acceleration of its internal diffusive motion upon supercoiling to intermediate density. Here, we present an independent study that shows a similar acceleration for fully supercoiled plasmid DNA. We have developed a method that allows fluorescent labeling of a 200-bp region, as well as efficient supercoiling by Escherichia coli gyrase. Compared to plain circular or linear DNA, the submicrosecond motion within the supercoiled molecules appears faster by up to an order of magnitude. The mean-square displacement as a function of time reveals an additional intermediate regime with a lowered scaling exponent compared to that of circular DNA. Although this unexpected behavior is not fully understood, it could be explained by conformational constraints of the DNA strand within the supercoiled topology in combination with an increased apparent persistence length.
Kalkbrenner
Thomas
T
FOM Institute for Atomic and Molecular Physics (AMOLF), Kruislaan, Amsterdam, The Netherlands.
Arnold
Axel
A
Tans
Sander J
SJ
D027081:DNA Gyrase / Q000378:metabolism; D004278:DNA, Superhelical / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics
D004278:DNA, Superhelical; D005456:Fluorescent Dyes; D027081:DNA Gyrase