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. Author manuscript; available in PMC: 2019 Dec 1.

Published in final edited form as: DNA Repair (Amst). 2018 Sep 20;72:56–63. doi: 10.1016/j.dnarep.2018.09.008

Fig. 3. — Single-turnover kinetics of SMUG1 activity in different sequence contexts. (A) Kinetic time courses were used to evaluate the excision of U in ssDNA substrates in 5'-UpT-3' (dashed line) and 5'-UpG -3' (solid line) sequence contexts. Reactions consisted of 20 nM DNA substrate, 640 nM SMUG1 in Buffer TNK. Data were fit to a single exponential equation. Error bars represent standard error (n=2 for 5'-UpT-3'; n=3 for 5'-UpG-3'). The observed rates and fraction product are k_obs=0.14 ± 0.01 min⁻¹, 0.92 (5'-UpT-3') and k_obs=0.02 ± 0.01 min⁻¹, 1.0 (5'-UpG-3'). (B) Kinetic time courses were used to evaluate the excision of U in duplex and NCP in a 5'-UpT/ApG-3' sequence context. Reactions consisted of 20 nM DNA substrate, 640 nM SMUG1 in buffer described in Materials and Methods. Data were fit to a single exponential equation (duplex), or double exponential equation (NCP). Error bars represent standard error (n=2 for duplex; n=6 NCP). Open squares (), duplex; filled squares (), NCP. The observed rates (fraction product) are k_obs=4.4 ±0.1 min⁻¹ (0.99) (duplex) and k_obs-fast=4.0 ± 0.9 min⁻¹ (0.06), k_obs-slow = 0.05 ± 0.02 min⁻¹¹ (0.14) (NCP).

Inline graphic — Single-turnover kinetics of SMUG1 activity in different sequence contexts. (A) Kinetic time courses were used to evaluate the excision of U in ssDNA substrates in 5'-UpT-3' (dashed line) and 5'-UpG -3' (solid line) sequence contexts. Reactions consisted of 20 nM DNA substrate, 640 nM SMUG1 in Buffer TNK. Data were fit to a single exponential equation. Error bars represent standard error (n=2 for 5'-UpT-3'; n=3 for 5'-UpG-3'). The observed rates and fraction product are k_obs=0.14 ± 0.01 min⁻¹, 0.92 (5'-UpT-3') and k_obs=0.02 ± 0.01 min⁻¹, 1.0 (5'-UpG-3'). (B) Kinetic time courses were used to evaluate the excision of U in duplex and NCP in a 5'-UpT/ApG-3' sequence context. Reactions consisted of 20 nM DNA substrate, 640 nM SMUG1 in buffer described in Materials and Methods. Data were fit to a single exponential equation (duplex), or double exponential equation (NCP). Error bars represent standard error (n=2 for duplex; n=6 NCP). Open squares (), duplex; filled squares (), NCP. The observed rates (fraction product) are k_obs=4.4 ±0.1 min⁻¹ (0.99) (duplex) and k_obs-fast=4.0 ± 0.9 min⁻¹ (0.06), k_obs-slow = 0.05 ± 0.02 min⁻¹¹ (0.14) (NCP).