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. 2003 Jan 1;17(1):77–87. doi: 10.1101/gad.1048303

Figure 1.

Figure 1

Replicative bypass of Tg by yeast DNA polymerases. (A) Structure of Tg formed upon oxidation of thymine. (B) DNA synthesis by yeast Polδ (1 nM), Polζ (1.5 nM), and Polη (1 nM) on undamaged and Tg-containing DNA substrates. (Lanes 1,2) Synthesis by Polδ on undamaged DNA (ND) and on DNA containing a Tg, respectively. (Lanes 3,4) Synthesis by Polζ on undamaged DNA and on DNA containing a Tg, respectively. (Lanes 5,6) Synthesis by Polη on undamaged DNA and on DNA containing a Tg, respectively. DNA length in nucleotides (nt) is given on the left. The position of the Tg is indicated by the arrow on the right. (C) Identification of the nucleotide inserted opposite Tg by Polδ (1 nM) and Polζ (1.5 nM). Synthesis was assayed on a DNA substrate containing a 19-nt primer annealed to either the undamaged or Tg-containing template. (Lane 1) A 19-nt primer. (Lanes 2,3) Synthesis by Polδ on undamaged DNA (ND) and on DNA containing a Tg, respectively. (Lanes 4,5) Synthesis by Polζ on undamaged DNA and on DNA containing a Tg, respectively. Lanes 6–9 show 20-nt markers containing the 19-nt primer sequence with an additional G, T, A, or C residue, respectively. For both B and C, reactions were carried out using the standard DNA polymerase assay containing 100 μM of each dNTP for 10 min at 30°C. The * in B and C indicates the position of the undamaged thymine or Tg in the template.