The N-Terminal Domain of Human DNA Helicase Rtel1 Contains a Redox Active Iron-Sulfur Cluster
Figure 5
DNA binding activity of RtelN with modified iron-sulfur cluster. (a) Effect of H2O2 on the RtelN iron-sulfur cluster. RtelN (20 μM) was incubated with the indicated concentrations of H2O2 (0 to 200 μM) at room temperature for 30 min. The UV-visible spectra were taken after incubation. (b) Effect of NO on the RtelN iron-sulfur cluster. RtelN (20 μM) was incubated with the indicated concentrations of the NO releasing reagent diethylamine NONOate (0 to 500 μM) at 37°C for 10 min. The UV-visible spectra were taken after incubation. (c) ssDNA binding activity of RtelN after the iron-sulfur cluster was modified. Untreated RtelN and RtelN treated with 200 μM H2O2 or 500 μM NONOate were incubated with the fluorescence-labeled ssDNA (0.5 μM). The DNA-protein complex and free DNA probe were resolved on a 0.6% agarose gel. The results are representative of three independent experiments.
