Effect of mutations in 3’ UTR on Core-binding and HCVtcp production.

(A) Schematic representation of designed mutants in replication-defective subgenomic replicon SGR-JFH1/Gluc/GND. Colored shadows and dashed lines were used to depict the deletion boundaries. (B) Production of HCVtcp by using the mutant subgenomic replicons. HCVtcp production (upper) and expression of subgenome (Gluc) and Core in the producer cells (middle and lower) were shown. (C) Predicted structures of SLI and II of 3’ UTR are depicted along with the substitution mutations introduced. The resultant mutants were named STIM, STIIM, LIM, LIIM and LI&IIM. (D) The interactions of Core with 3’ UTR mutants shown in (C). (E) Production of HCVtcp by replication-defective subgenomic replicons with 3’ UTR mutations shown in (C). HCVtcp production (upper), expression of subgenome and Core in the producer cells (middle and lower) are shown. HCVtcp production was determined as in Fig 2. Results shown represent the mean of three independent experiments ± SEM. HCV RNA copies are indicated as numbers per μg of total RNA for each assay, and Gluc activities are indicated as RLU per μl. VSL: variable region and poly (U/UC) tract, 3’X: the 3’ X tail, ×: GND mutants in NS5B.