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. 2004 Nov;78(21):11823–11832. doi: 10.1128/JVI.78.21.11823-11832.2004

FIG. 4.

FIG. 4.

Lysines 263, 280, and/or 284 are required for interaction with assembled proteasomes. 293T cells were transfected with the control pSG5 plasmid (lane 1) or with plasmids encoding Tax K1-10R (lane 2), Tax K4-10R (lane 3), Tax K6-8R (lane 4), Tax K1-3R (lane 5), and wild-type (wt) Tax (lane 6). (A) An aliquot of cell extract was subjected to direct immunoblot with patient sera (upper panel) and antiactin (lower panel). (B) Equal amounts of total proteins were immunoprecipitated with the antiproteasome antibody MCP21 and revealed by blotting with patient sera (upper panel) or an antibody specific for a proteasome β-subunit (lower panel). The positions of heavy chains (HC) and light chains (LC) revealed by the secondary antibody are indicated. (C) 293T cells were cotransfected with an HTLV-1 long terminal repeat-β-galactosidase reporter plasmid and a plasmid encoding wild-type or mutated Tax protein. The activity of the wild-type protein was arbitrarily set to 100% (wild-type induction = 18-fold).