Fig. 4: RAD52 is a m5C reader.
From: m5C modification of mRNA serves a DNA damage code to promote homologous recombination
a U2OS-TRE WT and TRDMT1 KO cells were transfected with TA-KR and stained for RAD51 1 h after light irradiation. Representative figures were shown (scale bar: 10 μm). b U2OS-TRE WT, TRDMT1 KO, and TRDMT1 stably rescued cell lines were transfected with TA-KR. and stained for RAD51 1 h after light irradiation. Fold increase of RAD51 foci intensity was calculated (n = 25, mean ± SD). c U2OS-TRE WT and TRDMT1 KO cells were transfected with TA-KR and GFP-RAD52 and fixed 1 h after light irradiation. Representative figures were shown (scale bar: 10 μm). d U2OS-TRE WT, TRDMT1 KO, and TRDMT1 stably rescued cell lines were co-transfected with GFP-RAD52 and TA-KR and quantified for RAD52 foci frequency (n = 3, mean ± SD). e RNA oligos with or without m5C modification and their complementary DNA oligos were chemically synthesized for RAD52 binding experiments. f RAD52 protein in the cell lysate was pulled down by biotin-labeled DNA:RNA hybrids, with or without m5C RNA modification, coated on streptavidin magnetic beads. g The binding of RAD52 protein to DNA:RNA hybrids with or without m5C RNA modification was measured in electrophoretic mobility shift assays and quantified (n = 3, mean ± SD). Statistical analysis was performed with the unpaired two tailed Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
