Figure 5.

Removal of 3′-PG group by the Rad1-Rad10 nuclease. 5′-End labeled DNA substrate PG1 or PG2 (50 fmole) with a 3′-PG modification (lanes 5-8,13-16) or unmodified (lanes 1-4,9-12) was incubated with equimolar amounts (50 fmole) of Rad1, Rad10, or Rad1-Rad10 protein complex for 30 min at 30°C. Lanes 1, 5, 9, and 13 contained DNA substrate alone in reaction buffer. Reaction products were analyzed by electrophoresis on a 10% sequencing gel with oligonucleotide size markers (indicated on the far right) and autoradiography. The oligomer carrying the 3′-PG modification or unmodified and labeled with ³²P at the 5′ end is indicated with an asterisk at the 3′ end.