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. 2022 Aug 5;298(9):102307. doi: 10.1016/j.jbc.2022.102307

Figure 5.

Figure 5

The SRAP DPC and DpC are reversible.A, reaction of YedK DPC with heat (Δ), followed by fresh YedK, AP-DNA, or buffer. An uncropped version of this gel is shown in Figure 5. B, time course of YedK and HMCES-SRAP DPC reversal at 37 °C, as measured by exchange of cross-linked DNAs of different lengths. DPC between YedK (top gel) or HMCES-SRAP (bottom gel) and 20-mer AP-DNA (DPC-20) was incubated with 4-fold excess 40-mer AP-DNA or buffer and analyzed at the indicated time points. Free protein and DPC were visualized via Coomassie stained SDS-PAGE. C, DPC reversal of YedK E105Q and H160Q mutants. D, spontaneous reversal of YedK DpC, preformed with 20-mer ssDNA and incubated with 4-fold excess aoN-g or buffer. DNA bands were visualized with FAM fluorescence. DPC, DNA–protein cross-link.