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Cyclization of DNA with sticky ends is commonly used to measure DNA bendability as a function of length and sequence, but how its kinetics depend on the rotational positioning of the sticky ends around the helical axis is less clear. Here, we measured cyclization (looping) and decyclization (unlooping) rates ( k  loop  and  k  unloop ) of DNA with sticky ends over three helical periods (100-130 bp) using single-molecule fluorescence resonance energy transfer (FRET).  k  loop  showed a nontrivial undulation as a function of DNA length whereas  k  unloop  showed a clear oscillation with a period close to the helical turn of DNA (∼10.5 bp). The oscillation of  k  unloop  was almost completely suppressed in the presence of gaps around the sticky ends. We explain these findings by modeling double-helical DNA as a twisted wormlike chain with a finite width, intrinsic curvature, and stacking interaction between the end base pairs. We also discuss technical issues for converting the FRET-based cyclization/decyclization rates to an equilibrium quantity known as the J factor that is widely used to characterize DNA bending mechanics.

Determinants of cyclization–decyclization kinetics of short DNA with sticky ends