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. 2012 Mar 14;109(13):5046–5051. doi: 10.1073/pnas.1200808109

Fig. 4.

Fig. 4.

DNA-specific endonuclease activity of CCHFV NP. (A) Time series of in vitro dsDNA (Upper) and ssDNA (Lower) and (B) dsRNA (Upper) and ssRNA (Lower) degradation assay products. Reaction products of 0.3 μM purified CCHFV NP with 100 ng/μL substrates at 37 °C in a final volume of 10 μL are shown after 5, 10, 20, 40, and 60 min. Reaction products were loaded onto a 20% (wt/vol) polyacrylamide gel and stained with ethidium bromide. (C) Effect of divalent cations on CCHFV NP nuclease activity. Purified CCHFV NP (0.3 μM) and dsDNA substrate were incubated with CoCl2, NiCl2, MgCl2, FeSO4, CaCl2, CuCl2, MnCl2, and ZnCl2 at a concentration of 1 mM. CCHFV NP alone, CCHFV NP and EDTA, or CCHFV NP and EDTA with Mn2+ were used as controls. All reactions were stopped by the addition of 10 mM EDTA. (D) Effect of CCHFV NP endonuclease activity on circular double-stranded plasmids. CCHFV NP was incubated with circular double-stranded plasmids for different lengths of time. All reactions were stopped by the addition of 10 mM EDTA.