TABLE 4.

Translation and replication of WT D2 and NS5 D4MT/D2POL chimera replicon RNAs

WT D2 Rluc replicon RNA and D2 replicon RNA encoding the D4MT/D2POL chimera with or without the K74I mutation in the D4MT/D2POL domain as well as with or without the D290N mutation within the D2 NS3 helicase domain were used in the experiments.

Sample	3 h	24 h	48 h	72 h
	RLUa
No RNA	577 ± 16	591 ± 48	582 ± 22	575 ± 9
WT D2 RNA	33,345 ± 360	2,451 ± 38	33,941 ± 471	205,219 ± 839
WT D2 RNA with NS3 D290N mutation	25,257 ± 295	3,347 ± 37	51,722 ± 1574	163,326 ± 1980
NS5 D4MT/D2POL chimera RNA	43,422 ± 673	1512 ± 4	1009 ± 4	1135 ± 50
NS5 D4MT/D2POL chimera RNA with NS3 D290N mutation	48,292 ± 503	1060 ± 24	744 ± 4	724 ± 16
NS5 D4MT K74I/D2POL chimera RNA	37,600 ± 333	942 ± 42	857 ± 52	862 ± 29
WT-noncapped D2 replicon RNA	541 ± 35	496 ± 11	442 ± 5	762 ± 21

^a The Rluc (RLU) values are expressed as mean with S.D. from duplicate wells. Fluc RNA was cotransfected as an internal control to monitor the transfection efficiencies as described under “Experimental Procedures.” The RLU values were normalized in each case using the Fluc activities measured at 3 h time points from the cotransfection experiments of Fluc RNA. Fluc activities of samples are shown in Table 2.