FIGURE 7.

A, purified recombinant SsOGT was incubated for 1 h at 37 °C with 4-fold molar excess of either ds-UP or ds-UP^m oligonucleotides, to obtain SsOGT or the methylated form (SsOGT^m), respectively. 50 or 100 ng of either SsOGT or SsOGT^m was mixed and incubated for 30 min at 70 °C with extracts from control or 0.35 mm MMS-treated cultures (200 μg/lane). Filters were probed with the anti-His-tag antibody. B, soluble extracts (80 μg/lane) from control cultures were incubated for 1 h at 70 °C in the presence of MMS (1.0 mm) and the ds-UP^m or ds-UP oligonucleotides. R, 200 ng of purified SsOGT. Filters were stripped and probed sequentially with anti-SsOGT (to follow the endogenous SsOGT) and anti-β-prefoldin (a-βpref.) (to control for protein loading).