FIG. 7.
Inhibition activities and RNAP II interactions of RS domain mutants. (A) Various T-RS domain deletion mutants were tested for dominant negative inhibition on an HIV-1 LTR-RREIIB-FFL reporter, and the values shown represent percent inhibition at a 1:1 activator/inhibitor plasmid ratio relative to activation without inhibitor. The data are mean values from three independent experiments. (B) Sequences of RS dipeptide substitutions and their inhibition activities with a T-Rev activator (white bar) and HIV-1 LTR-RREIIB-FFL reporter at 1:0.2 (black bars) and 1:1 (gray bars) ratios of activator to inhibitor. (C) HeLa cells were transfected with HA-tagged versions of the T-RS25-63 domain or mutants (expression levels shown on the left top panel) with loading controls (levels shown on left bottom panel). Amounts of associated RNAP IIa were assessed by immunoprecipitation (IP) with anti-HA (α-HA) and Western blotting with the 8WG16 (α-8WG16) antibody (right panel).