FIG. 10.

P1 nuclease assay showing binding of RAGs to the single-stranded region of the symmetric bubble. Increasing concentrations of P1 nuclease were incubated with a [γ-³²P]ATP end-labeled 50-bp ds oligomers containing a symmetric bubble structure, with and without pretreatment with RAGs (100 ng of RAGs treated for 10 min at 37°C, in a volume of 15 μl) in buffer A for 15 min for 37°C. The reaction products were then resolved either on a 15% native polyacrylamide gel (A) or a 15% denaturing polyacrylamide gel (B). In both panels, lanes 1 to 5 contain no RAGs and lanes 6 to 10 are with RAGs. The shifted band due to the nicked bubble is indicated by an arrow in panel A. Lane M, 50-bp ladder; lane M′, 22-nt oligomer digested with Klenow to generate a ladder; lane O, [γ-³²P]ATP-labeled 22-nt oligomer. The bubble region is indicated by the bracket.