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. Author manuscript; available in PMC: 2017 May 7.
Published in final edited form as: Nat Struct Mol Biol. 2016 Nov 7;23(12):1092–1100. doi: 10.1038/nsmb.3319

Figure 1.

Figure 1

8-oxodGTP is a telomerase chain terminator. (a) Telomerase catalytic cycle. Blue indicates newly added nucleotides and grey indicates the telomerase template. Numbers represents steps in the cycle. (b) Telomerase reactions were conducted using (TTAGGG)3 primer (3R) with high dNTPs (500 μM dGTP, 500 μM dATP, and 2.9 μM dTTP) (lanes 2 – 4) or cellular dNTPs (37 μM dTTP, 24 μM dATP, 29 μM dCTP, 5.2 μM dGTP) (lanes 6 – 8) and 0.3 μM 32P-α-dTTP. dGTP and 8-oxodGTP were mixed at 1:0, 1:1, and 0:1 ratios for a total of 500 μM or 5.2 μM unlabeled guanine nucleotide. Products were separated on denaturing gels. The LC was a 32P end-labeled 18-mer oligonucleotide. Numbers on the left show number of added repeats, and letters on right indicate template base. (c) Total products were normalized to the LC and used to calculate processivity and relative activity. Bars represent the mean ± sd from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, two-tailed Student’s t test. N.D. = not determined. Uncropped gel image is shown in Supplemental Data Set 1.