Fig. 2
From: RAD18 methylation by the methyltransferase SETD6 attenuates DNA breaks
SETD6 methylates RAD18 on K73 and K406. (A) In-vitro methylation assay in the presence of 3H-labeled SAM with recombinant His-SETD6 and His-RAD18. The methylated proteins were detected by autoradiogram and a Coomassie stain of the recombinant proteins used in the reactions is shown on the bottom. (B) Semi-in-vitro methylation assay of immunoprecipitated Flag-RAD18 that was over-expressed in PC3 cells, with and without the presence of His-SETD6. The methylated proteins were detected by autoradiogram and Flag-RAD18 was visualized using Western blot analysis using αFlag antibody. (C) HeLa cells lysates were immunoprecipitated with Pan-Methyl lysine antibody followed by Western Blot analysis with the indicated antibodies. (D) MS spectra of RAD18 TQCPTCCVTVTEPDLK73NNR and LSSVCMGQEDNMTSVTNHFSQSK406. Major y- and b-ions are displayed as red, and blue, respectively. (E) In-vitro methylation assay in the presence of 3H-labeled SAM with the indicated recombinant proteins. Coomassie stain of the recombinant proteins used in the reactions is shown on the bottom.
