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Figure 3 | Retrovirology

Figure 3

From: The histone chaperone protein Nucleosome Assembly Protein-1 (hNAP-1) binds HIV-1 Tat and promotes viral transcription

Figure 3

Mapping of hNAP-1 and Tat interacting domains. A. Schematic representation of hNAP-1 protein and of its deletion mutants obtained as GST fusion proteins. The capacity of binding to Tat – see experiment in panel B – is indicated on the right side of each mutant. The two dotted boxes indicate the hNAP-1 domains interacting with Tat. B. Representative GST pulldown experiment using the indicated hNAP-1 mutants and radiolabelled Tat101 protein. The autoradiography shows the amount of Tat binding to each mutant; the histogram on top shows densitometric quantification of data, expressed as fold binding with respect to background binding to GST alone (set as 1). The lower panel shows the Coomassie stained gel at the end of the binding experiment. The experiment was repeated at least three times with similar results. C. Schematic representation of HIV-1 Tat protein and of its mutants obtained as GST fusion proteins. The capacity of binding to hNAP-1 – see experiment in panel D – is indicated on the right side of each mutant. The dotted box corresponds to the basic domain of Tat, which binds hNAP-1. D. Representative GST pulldown experiment using the indicated Tat mutants (obtained as GST fusion proteins) and in vitro transcribed and translated hNAP-1 protein. The autoradiography shows the amount of hNAP-1 binding to each mutant; the histogram on top shows densitometric quantification of data, expressed as fold binding with respect to background binding to GST alone (set as 1). The lower panel shows the Coomassie stained gel at the end of the binding experiment. The experiment was repeated at least three times with similar results.

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