FIG. 3.

GalT is closely associated by coimmunoprecipitation with the KUN nonstructural proteins NS5, NS3, NS1, NS2A, and NS4A, which are proposed to constitute the flavivirus replication complex (29, 49). Infected Vero cells were radiolabelled (in the presence of actinomycin D) from 24 to 28 h p.i. and harvested in RIP assay buffer for RIP with either anti-NS3 (lanes 3 and 4) or anti-GalT (lanes 5 and 6) antibodies. Lanes 1 and 2 represent the initial lysates before RIP. Specific KUN nonstructural proteins are indicated by arrows, and the KUN structural proteins E, prM, and C (notably absent in the immunoprecipitate in lane 6) are indicated by asterisks in lane 2. Lanes M, mock-infected cell lysates; lanes K, KUN-infected cell lysates. Proteins were separated on an SDS–12.5% polyacrylamide gel and visualized by autoradiography. Sizes of protein markers, indicated on the left, are expressed in kilodaltons.