Fig. 3: PRMT3 increases H4R3Me2a levels and chromatin accessibility at HIV-1 LTR to promote Tat-dependent recruitment of P-TEFb to the LTR. | Nature Communications

Fig. 3: PRMT3 increases H4R3Me2a levels and chromatin accessibility at HIV-1 LTR to promote Tat-dependent recruitment of P-TEFb to the LTR.

From: PRMT3 reverses HIV-1 latency by increasing chromatin accessibility to form a TEAD4-P-TEFb-containing transcriptional hub

Fig. 3

a, b H4R3Me2a levels were examined in WT and PRMT3 KO cells (a), or in indicated cells treated with DMSO or SGC707 (b). Densitometric analyses of immunoblots are shown. c, d H4R3Me2a occupancy at LTR-luciferase regions in WT and KO cells (c) (p = 0.0050, p = 0.0133, p = 0.0102, p = 0.0047), and cells treated with DMSO or SGC707 (d) (p = 0.0064, p = 0.0009, p = 0.0256, p = 0.1042) were measured. e, f ATAC-Seq was performed in WT and KO cells transfected with Tat using three biological replicates (e). Normalize depth (Reads per million) was calculated using LTR counts divided by the number of sequences to HIV-1 × 1,000,000. The center line, upper, and lower edges of the boxplot represented the mean, first, and third quartile (f). g–i WT and KO cells were cotransfected with Tat and Flag-CycT1 (g) (p = 0.0029, p = 0.0028, p = 0.0109, p = 0.0019) or Flag-CDK9 (h) (p = 0.0050, p = 0.0042, p = 0.0135, p = 0.0183). F1C2 cells were treated with DMSO or SGC707 (i) (p = 0.0007, p = 0.0035, p = 0.0014, p = 0.00009). The occupancy of Flag-CycT1 or Flag-CDK9 at LTR-luciferase regions was measured. j–l Luciferase activities were measured for Tat transfected (j) (p = 0.0017, p = 0.0158), JQ1 (k) (p = 0.0005, p = 0.0065), or PMA (l) (p = 0.000003, p = 0.0011) treated WT and KO cells when CycT1 overexpression. m–q Luciferase activities were measured for Tat transfected (m) (p = 0.0005, p = 0.0041), JQ1 (n) (p = 0.0033, p = 0.0137), or PMA (o) (p = 0.0042, p = 0.0019) treated cells overexpression CycT1 when treated with SGC707, in WT, CDK9 KD cells (p = 0.0147, p = 0.0099, p = 0.0270) (p), or in NH1 cells transfected with indicated plasmid together with Tat (q) (p = 0.0008, p = 0.0025, p = 0.0003). Error bars = mean +/− SD of three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. denotes no significance, two-tailed t test except (f) was assessed by Wilcoxon’s test. Source data are provided as a Source Data file.

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