“By regulating access to the genetic
code, DNA supercoiling strongly affects DNA metabolism. Despite its
importance, however, much about supercoiled DNA (positively
supercoiled DNA, in particular) remains unknown. Here we use electron
cryo-tomography together with biochemical analyses to investigate
structures of individual purified DNA minicircle topoisomers with
defined degrees of supercoiling. Our results reveal that each
topoisomer, negative or positive, adopts a unique and surprisingly
wide distribution of three-dimensional conformations. Moreover, we
uncover striking differences in how the topoisomers handle torsional
stress. As negative supercoiling increases, bases are increasingly
exposed. Beyond a sharp supercoiling threshold, we also detect
exposed bases in positively supercoiled DNA. Molecular dynamics
simulations independently confirm the conformational heterogeneity
and provide atomistic insight into the flexibility of supercoiled
DNA. Our integrated approach reveals the three-dimensional structures
of DNA that are essential for its function. ” -abstract from Nature
Using a combination of approaches
supercoiling causes DNA to adopt large conformational variability.
Unpaired bases and localized distortions contribute to this
variability both by relieving torsional strain and by providing
flexible hinges. Positive and negative supercoiling are accommodated
differently.
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