Table 5.

Phenotypic analysis of JFH-1 GT 2a replicon variants^a

Change and mutation	Fold change in EC50	Fitness (% relative to WT)
Single-residue changes
T179A	1.1 ± 0.2	89.0 ± 11.2
S282T	2.8 ± 0.7	30.5 ± 8.1
M289L	2.9 ± 0.9	99.9 ± 9.8
I293L	1.3 ± 0.3	93.9 ± 5.4
M434T	0.9 ± 0.2	104.7 ± 13.7
H479P	1.0 ± 0.3	126.0 ± 37.1
Combination of residue changes in finger and palm domains
T179A/M289L	2.1 ± 0.2	44.8 ± 10.6
M289L/I293L	2.5 ± 0.5	150.4 ± 16.7
T179A/M289L/I293L	3.5 ± 0.6	95.6 ± 26
S282T/M289L	6.3 ± 1.0	32.3 ± 4.9
S282T/M289L/I293L	5.6 ± 1.9	45.2 ± 9.3
T179A/S282T/I293L	8.3 ± 0.7	1.8 ± 0.6
T179A/S282T/M289L/I293L	9.5 ± 2.5	6.6 ± 2.2
Combination of residue changes in finger, palm, and thumb domains
S282T/M289L/H479P	7.4 ± 0.6	72.5 ± 12.6
S282T/M289L/I293L/M434T	7.1 ± 0.1	83.0 ± 9.3
S282T/M289L/I293L/H479P	8.4 ± 0.5	126.0 ± 6.9
T179A/S282T/I293L/H479P	9.0 ± 1.5	85.5 ± 11.3
T179A/S282T/M289L/I293L/M434T	10.6 ± 1.9	45.1 ± 6.5
T179A/S282T/M289L/I293L/H479P	11.8 ± 3.0	38.8 ± 5.3
T179A/S282T/M289L/H479P	11.4 ± 1.2	0.26 ± 0.13

^aMutations in JFH-1 NS5B were generated by mutagenesis, and replicons were electroporated into Lunet cells, which were subjected to G418 selection prior to the susceptibility studies. Changes within the finger domain (T179A and S282T) are in bold, of which S282T is also underlined. Changes within the palm domain (M289L and I293L) are in italic. Changes within the thumb domain (M434T and H479P) are normal text. EC₅₀ fold change was determined by normalizing the EC₅₀s for PSI-7977 from the replicon variants with that of the wild type (WT). Replication fitness was determined by measuring the expression of luciferase at 4 h and 96 h posttransfection and normalizing the luminescence levels with that of the wild type. All values are reported as average ± standard deviation from at least three independent experiments performed in duplicate.