Fig. 5. Effect of incubation temperature on the inhibitory effect of CL on ORC binding to origin DNA. ORC (0.15 pmol) was pre-incubated with the indicated amounts of CL for 10 min at 4 or 30°C and further incubated with radio-labeled ARS1 DNA fragments (90 fmol) at 4 or 30°C. The amount of bound DNA was determined by a filter-binding assay. In the absence of CL, 4.1 or 6.0 fmol of DNA was bound to ORC at 4 or 30°C, respectively. Values are mean ± SD, n = 3.