Timing Facilitated Site Transfer of an Enzyme on DNA

Many enzymes that react with specific sites in DNA exhibit the property of facilitated diffusion, where the DNA chain is used as a conduit to accelerate site location. Despite the importance of such mechanisms in gene regulation and DNA repair, there have been few viable approaches to elucidate the microscopic process of facilitated diffusion. Here we describe a new method where a small molecule trap (uracil) is used to clock a DNA repair enzyme as it hops and slides between damaged sites in DNA. The “molecular clock” provides unprecedented information: the mean length for DNA sliding, the 1D sliding constant, the maximum hopping radius and time frame for DNA hopping events. In addition, the data establish that the DNA phosphate backbone is a sufficient requirement for DNA sliding.