Figure 4
Region of bovine lentiviral Vif responsible for degradation of gaur A3Z3.
(A) Alignment of the amino acid sequences of BIV and JDV Vif. Asterisks indicate the identical amino acid. The determined responsible region is indicated by a blue line (residues 57–102; identical to that in panel C). The S/TLQ motif is indicated by yellow shading (residues 150–152). (B) Western blotting. The HA-tagged expression plasmid for cattle A3Z3 (150 ng) was co-transfected with the flag-tagged expression plasmids for wild-type (WT) or SLQ/AAA-mutated BIV/JDV Vif (1,000 ng). (C,D) Exchanging derivatives of BIV and JDV Vif. (C) Scheme of a series of exchanging mutants. The names of respective derivatives (indicated on the left of the panel) are identical to those in panel D. The determined responsible region is indicated by a blue line (residues 57–102; identical to that in panel A). (D) Western blotting. The HA-tagged expression plasmids for cattle or gaur A3Z3 (150 ng) were co-transfected with the flag-tagged expression plasmids for a series of BIV/JDV Vif derivatives (1,000 ng). In panels B and D, the value under the band of each bovine A3Z3-HA represents the percentage of band intensity compared to the absence of Vif. Blots have been cropped; uncropped blots are shown in Supplementary Figure 8.
