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Figure 4 | Retrovirology

Figure 4

From: GCN5-dependent acetylation of HIV-1 integrase enhances viral integration

Figure 4

GCN5-mediated acetylation increases the catalytic activity of IN. (A) Schematic representation of IN-GCN5 tethered catalysis constructs. Full-length IN, tagged with a N-terminal 6× His epitope, is fused in frame with TEV proteolytic site and cloned upstream of the 6-300 amino acid region of wild type GCN5 (IN-HAT wt) or its catalytically inactive allele (IN-HAT mut). (B) 1 μg and 2 μg of IN derived from IN-HAT wt (lanes 1 and 3, respectively), or 1 μg and 2 μg of IN derived from IN-HAT mut (lanes 2 and 4, respectively) were analyzed by Western blotting with anti-acetyl-lysine antibody (top panel) or anti-IN antibody (bottom panel). (C) 3' -end processing activity of IN derived from IN-HAT wt (lane 1: 100 ng; lane 3: 200 ng) or IN-HAT mut (lane 2: 100 ng; lane 4: 200 ng). Lane 5: DNA substrate; lane 6: DNA substrate with 40 ng of 6× His-tagged IN. (D) Strand transfer activity of IN derived from IN-HAT wt (lane 1: 100 ng; lane 3: 200 ng) or IN-HAT mut (lane 2: 100 ng; lane 4: 200 ng). Lane 5: DNA substrate; lane 6: DNA substrate with 40 ng of 6× His-tagged IN. In (C) and (D), the DNA substrate (S) and the catalytic products (P) are indicated.

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