DNA Bending Force Facilitates Z-DNA Formation under Physiological Salt Conditions

[Image: see text] Z-DNA, a noncanonical helical structure of double-stranded DNA (dsDNA), plays pivotal roles in various biological processes, including transcription regulation. Mechanical stresses on dsDNA, such as twisting and stretching, help to form Z-DNA. However, the effect of DNA bending, one of the most common dsDNA deformations, on Z-DNA formation is utterly unknown. Here, we show that DNA bending induces the formation of Z-DNA, that is, more Z-DNA is formed as the bending force becomes stronger. We regulated the bending force on dsDNA by using D-shaped DNA nanostructures. The B–Z transition was observed by single-molecule fluorescence resonance energy transfer. We found that as the bending force became stronger, Z-DNA was formed at lower Mg(2+) concentrations. When dsDNA contained cytosine methylations, the B–Z transition occurred at 78 mM Mg(2+) (midpoint) in the absence of the bending force. However, the B–Z transition occurred at a 28-fold lower Mg(2+) concentration (2.8 mM) in the presence of the bending force. Monte Carlo simulation suggested that the B–Z transition stabilizes the bent form via the formation of the B–Z junction with base extrusion, which effectively releases the bending stress on DNA. Our results clearly show that the bending force facilitates the B–Z transition under physiological salt conditions.