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. 2018 Jun 18;46(14):7169–7178. doi: 10.1093/nar/gky525

Figure 5.

Figure 5.

Effect of ssDNA overhang polarity on RPA-dependent and RPA-independent uracil excision selectivity. (A) Junction substrate with a 32 nt 3′ ssDNA overhang. The polarity of this substrate is reversed compared to those used in Figures 1–3, but the uracils were maintained 9 and 33 bp from the ssDNA–dsDNA junction. (B) Reaction of 900 pM hUNG2 with the 3′ ssDNA overhang junction substrate in panel A in the presence and absence of RPA. A reaction was also performed in the absence of RPA with an identical duplex substrate that had no overhang. For all assays, DNA concentration was 3 μM. (C) Selectivity of hUNG2 for U9 and U33 in the presence and absence of RPA, and also in the absence of RPA without an overhang on the duplex. The selectivity was calculated from the band intensities in panel B.