Novel ATP-Independent RNA Annealing Activity of the Dengue Virus NS3 Helicase
Figure 2
NTP utilization in RNA unwinding reactions.
(A) Different nucleotides were tested as substrates for the RNA unwinding activity of NS3-hel. Unwinding reactions were carried out at 30°C for 30 minutes as described in Fig. 1E including 1.0 mM of the respective nucleotide as indicated on top of each lane. A negative control (no nucleotide) was included. (B) Usage of canonical NTPs for the RNA unwinding activity of NS3-FL and NS3-hel. Unwinding reactions were carried out as described in Fig. 1E, but incubated at 37°C in the presence of 2.0 mM of ATP, CTP, GTP or UTP as indicated on the right. Lane H represents a positive control (heat denatured duplex). Negative control (no NS3), NS3-hel, or NS3-FL were included as indicated on top of the gel. Mobility of double-stranded (dsRNA) and single-stranded RNA probe (ssRNA) is indicated on the left.