HIV-1 persists lifelong in memory cells of the immune system as latent provirus that rebounds upon treatment interruption. Therefore, the latent reservoir is the main target for an HIV cure. Here, we studied the direct link between integration site and transcription using LEDGINs and Barcoded HIV-ensembles (B-HIVE). LEDGINs are antivirals that inhibit the interaction between HIV-1 integrase and the chromatin-tethering factor LEDGF/p75. They were used as a tool to retarget integration, while the effect on HIV expression was measured with B-HIVE. B-HIVE tracks insert-specific HIV expression by tagging a unique barcode in the HIV genome. We confirmed that LEDGINs retarget integration out of gene-dense and actively transcribed regions. The distance to H3K36me3, the marker recognized by LEDGF/p75, clearly increased. LEDGIN treatment reduced viral RNA expression and increased the proportion of silent provirus. Finally, silent proviruses obtained after LEDGIN treatment were located further away from epigenetic marks associated with active transcription. Interestingly, proximity to enhancers stimulated transcription irrespective of LEDGIN treatment, while the distance to H3K36me3 only changed after treatment with LEDGINs. The fact that proximity to these markers are associated with RNA expression support the direct link between provirus integration site and viral expression.

中文翻译：

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HIV-1前病毒整合位点的染色质景观决定了病毒的表达。

HIV-1作为潜在的原病毒在免疫系统的记忆细胞中持续存在，在治疗中断后会反弹。因此，潜在的水库是治疗HIV的主要目标。在这里，我们研究了使用LEDGIN和条形码HIV集成体（B-HIVE）整合位点与转录之间的直接联系。LEDGIN是抑制HIV-1整合酶与染色质束缚因子LEDGF / p75之间相互作用的抗病毒药物。它们被用作重新整合的工具，而B-HIVE可以测量对HIV表达的影响。B-HIVE通过标记HIV基因组中唯一的条形码来跟踪特定于HIV的表达。我们证实，LEDGINs重新定位了基因密集和活跃转录区域之外的整合。到LEDGF / p75识别的标记物H3K36me3的距离明显增加。LEDGIN治疗降低了病毒RNA的表达并增加了沉默前病毒的比例。最后，LEDGIN处理后获得的沉默原病毒位于远离与主动转录相关的表观遗传标记的位置。有趣的是，与LEDGIN的治疗无关，与增强子的接近会刺激转录，而到H3K36me3的距离仅在使用LEDGIN进行治疗后才发生改变。这些标记的接近性与RNA表达有关的事实支持前病毒整合位点与病毒表达之间的直接联系。不管是否使用LEDGIN，与增强子的接近都会刺激转录，而仅使用LEDGIN治疗后，与H3K36me3的距离才会改变。这些标记的接近性与RNA表达有关的事实支持前病毒整合位点与病毒表达之间的直接联系。不管是否使用LEDGIN，与增强子的接近都会刺激转录，而仅使用LEDGIN治疗后，与H3K36me3的距离才会改变。这些标记的接近性与RNA表达有关的事实支持前病毒整合位点与病毒表达之间的直接联系。