FIG. 2.

Characterization of minimal requirements for NS5A hyperphosphorylation. Hep3B cells were transfected with the indicated constructs and labelled with either [³⁵S]methionine (A and B) or [³²P]orthophosphate (C) for 3 h. NS5A was immunoprecipitated with NS5A antiserum, and protein was loaded onto an SDS–7.5% polyacrylamide gel. (B and C) Immunoprecipitated proteins were incubated with (+) or without (−) λ-phosphatase as described in Materials and Methods prior to loading for SDS-PAGE. NS5A p56 and p58 are indicated by arrows on the right; the sizes of molecular weight marker proteins (M) are indicated on the left. BK, HCV proteins derived from the BK strain; H, HCV proteins derived from the H strain.