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• W2954141106 abstract "Abstract In Cre site-specific recombination, the synaptic intermediate is a recombinase homotetramer containing a pair of DNA target sites. The strand-exchange mechanism proceeds via a Holliday-junction (HJ) intermediate; however, the geometry of the DNA segments in the synapse has remained highly controversial. In particular, all crystallographic structures are consistent with an achiral planar Holliday-junction (HJ) structure, whereas topological assays based on Cre-mediated knotting of plasmid DNAs are consistent with a right-handed chiral junction. Here we use the kinetics of loop closure involving closely spaced (131-151 bp), directly repeated loxP sites to investigate the in-aqueo ensemble of conformations for the longest-lived looped DNA intermediate. Fitting the experimental site-spacing dependence of the loop-closure probability, J , to a statistical-mechanical theory of DNA looping provides evidence for substantial out-ofplane HJ distortion. This result unequivocally stands in contrast to the square-planar intermediate geometry determined from crystallographic data for the Cre-loxP system and other int-superfamily recombinases. J measurements carried out with an isomerization-deficient Cre mutant suggest that the apparent geometry of the wild-type complex may result from the temporal averaging of diverse right-handed and achiral structures. Applied to Cre recombinase, and other biological systems, our approach bridges the static pictures provided by crystal structures and the natural dynamics of macromolecules in vivo . This approach thus advances a more comprehensive dynamic analysis of large nucleoprotein structures and their mechanisms." @default.
• W2954141106 created "2019-07-12" @default.
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• W2954141106 date "2019-07-06" @default.
• W2954141106 modified "2023-09-27" @default.
• W2954141106 title "Loop-closure Kinetics Reveal a Stable, Right-handed DNA Intermediate in Cre Recombination" @default.
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• W2954141106 doi "https://doi.org/10.1101/695130" @default.
• W2954141106 hasPublicationYear "2019" @default.
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