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. Author manuscript; available in PMC: 2010 Nov 1.
Published in final edited form as: J Neurochem. 2008 Sep 20;107(3):734–744. doi: 10.1111/j.1471-4159.2008.05644.x

Figure 5. Processing of gapped DNA with a 5′-THF terminus in brain and mucosal extracts supplemented with purified repair proteins.

Figure 5

Gapped 5′-THF substrate was incubated with extracts alone or with extracts supplemented with indicated purified repair proteins. Reaction aliquots were taken at different time points and resolved in denaturing polyacrylamide gels. Products generated after 15- and 45-minute incubations are shown. With brain extracts, the 45-mer was observed after 45 but not 15 minutes (lane 2, bullet). In contrast, with mucosa, the 45-mer was transiently seen at 15 but not 45 minutes (lane 8, bullet). With mucosa, a larger product appeared at 15 minutes and then persisted as a sole product at 45 minutes (lane 8, asterisk). With mucosa, nucleolytic activities cause some degradation of the 45-mer at longer incubation times (lane 11). Purified proteins were added as indicated; in the brain addition of pol β, ligases, PCNA or FEN1 stimulated generation of the 45-mer in time dependent manner (lanes 3–7). In the case of FEN1, generation of a [+2] and longer extension products was strongly stimulated (lane 7). In contrast, supplementation of pol-β or FEN1 (lanes 8–9) had no significant effect on levels of products generated by mucosal extracts.