Genome-wide study of human methylation profile of J-Lat cells upon EGCG-treatment and UHRF1 knockdown

HIV-1 latent reservoirs constitute a major barrier to viral eradication. The various potencies of latency-reversing agents reflect the multiplicity of the silencing mechanisms underlying latency, and the dynamic and heterogeneous nature of the HIV-1 latent reservoirs. Here, we investigated the mechanisms of the DNA methylation inhibitor 5-aza-2’-deoxycytidine (5-AzadC) potency in HIV-1 latency reversal. We report that this potency was directly linked to 5-AzadC-induced demethylation of the HIV-1 promoter, which occurred at non-random CpG dinucleotides. Specifically, the methylation status of one of those CpGs allowed the recruitment of the epigenetic integrator UHRF1 to the HIV-1 promoter. 5-AzadC-induced viral reactivation led to a decreased in vivo recruitment of UHRF1. RNA interference-mediated knockdown of UHRF1 demonstrated its role in DNA methylation-mediated silencing of the latent HIV-1 promoter. Pharmacological downregulation of UHRF1 in ex vivo HIV+ patient cell cultures resulted in potent reactivation of latent HIV-1, further highlighting that UHRF1 could constitute a new molecular target to devise innovative HIV-1 curative strategies. Total genomic DNA from J-Lat cells. EGCG-treated cells, UHRF1 knock down and their respectives controls were analysed.