FIGURE 5.

Standing start assays for replication bypass of DNA-protein cross-links by hPol κ. 9-mer FAMdT primers were annealed with 20-mers containing unmodified dG (A), deaza-DHP-dG (B), or covalent cross-links to 10-mer peptide EQKLISEEDL (C), 23-mer peptide PDAQLVPGINGKAIHLVNNESSE (D), or full-sized protein AlkB (E) or histone (His) H4 (F). The resulting primer-template duplexes (0.15 μm) were incubated in the presence of hPol κ (0.30 μm). Reactions were initiated by the addition of a mixture of dNTPs (500 μm) and quenched at the indicated time points. Extension products were separated by 20% denaturing PAGE and visualized by fluorescence imaging.