Protein Kinase D3 Is Essential for Prostratin-Activated Transcription of Integrated HIV-1 Provirus Promoter via NF-κB Signaling Pathway
Figure 1
Prostratin enhances the transcription initiation of latent HIV-1 provirus. (a) Time course of prostratin-induced HIV-1 expression. HeLa cells with an integrated HIV-LTR-luciferase reporter gene (HIV-LTR-Luc) were treated with 2 μM of prostratin for 0–6 hrs, and the cell lysates were prepared for the detection of luciferase activity. The expression levels of luciferase gene were plotted based on 3 independent experiments, with the level of untreated cells set to 1.0. (b) Titration assay of prostratin-induced transcription activation of latent HIV-1 provirus. 2D10 cells, a Jurkat-based cell line containing the latent HIV-1 provirus with eGFP in place of Nef, were treated with indicated concentration of prostratin for 16 hrs. The GFP-positive cells were measured by flow cytometry and plotted as percentage of total cells based on 3 independent experiments. (c) Effect of prostratin treatment on transcription initiation and elongation of HIV-1. The schematics of the HIV-LTR-luciferase reporter gene (HIV-LTR-Luc) and the locations of qRT-PCR primers for detecting transcripts corresponding to the region of initiation (+1~+59, TAR) or elongation (+496~593, luciferase) are illustrated in top panel. HeLa cells with integrated HIV-LTR-luciferase gene were treated with 2 μM of prostratin for 4 hrs. The transcription levels were detected by qRT-PCR and plotted based on 3 independent experiments, with the level of untreated cells set to 1.0.
