Skip to main content
Figure 4 | Virology Journal

Figure 4

From: Phosphorylation of HIV Tat by PKR increases interaction with TAR RNA and enhances transcription

Figure 4

PKR phosphorylation of HIV-1 Tat wild-type and mutants. HIV-1 Tat wild-type and mutant proteins were expressed in BL21(DE3)pLysS cells from pET-DEST-42 expression clones, and purified by passage through a TALON™ cobalt affinity resin. PKR was immunoprecipitated from HeLa cell extracts, and activated with dsRNA in the presence of ATP. The phosphorylation reactions contained 2 μg of Tat protein, 6 μL of activated PKR suspension, and DBGA to a final volume of 12 μL. Phosphorylation was preformed at 30°C for the times indicated, in the presence of 2 μCi of γ-32P-ATP. Protein samples were analyzed by 15% SDS-PAGE. This figure only shows one representative gel out of three separate phosphorylation experiments performed for each protein.

Back to article page