FIG. 5.

RTA K152 E failed to efficiently bind to ISREs or RREs. A. RTA K152 E binds to ISRE with reduced affinities. Equal amounts of partially purified recombinant RTA and RTA K152E proteins from bacteria were used for EMSA with various probes. The probes were ISRE from the ISG-15 gene, vIL6 ISRE1, and K14 ISRE. B. RTA K152 E binds to RREs with reduced affinities. Two known RREs, PAN and ORF57 RREs, were used for EMSA. Cold competitors and protein used are indicated on top. The RTA-DNA complexes are shown. C. Coomassie blue staining of the partially purified E. coli-derived RTA and its mutant RTA K152E proteins. Molecular mass (MW) markers and their sizes in kilodaltons are also shown. D. Western blot analysis of E. coli-derived RTA and its mutant RTA K152E. His tag and RTA antibodies were used for detection of partially purified proteins as shown in panel C. Some MW markers were also reactive to His tag antibody as shown.