Studies on Epigenetically Active Latent Chromatin Maintenance

通过调节元件的突变或负责任的突变来解除对基因组相互作用的管制 细胞核酶导致多种疾病。尽管适当的基因组-基因组的意义 相互作用已经从疾病表型中看到，这是基因组环路的潜在分子机制 调控和转录的直接结果仍然没有很好的联系。这在一定程度上源于 细胞启动子调控的复杂性，因为它受多个转录因子和增强子控制 在细胞群体中具有显著噪声的元素。 重组卡波西肉瘤相关的转录组学、基因组学和蛋白质组学研究 疱疹病毒(KSHV)异构体鉴定宿主细胞ChAHP(CHD4，ADNP，HP1)蛋白复合体形成 与KSHV潜伏期相关核抗原(LANA)的稳定蛋白复合体在维持 潜伏病毒染色质的诱导性。已知ChAHP复合体限制细胞增强子的可及性和 调节细胞谱系，尽管蛋白质复合体如何调节增强子-启动子相互作用尚不清楚。 在这里，我们假设KSHV潜伏期裂解开关是增强子-启动子相互作用的产物，受 LANA-ChAHP复合体。在本应用程序中，我们将研究KSHV Episome是如何保持为诱导性的 如果我们能以蛋白质复合体为靶点进行治疗干预。细胞因子的分子作用 LANA/ChAHP蛋白质复合体将从三个不同的角度进行研究：(I)生化、(Ii)遗传和(Iii) 蛋白质复合体结构。通过利用已定义的增强子-启动子对和方便的可诱导mini 病毒染色质，我们将通过解剖ChAHP蛋白复合体功能来研究增强子-启动子的调控。这个 建议的研究不仅应该有助于理解KSHV潜伏期-裂解开关，而且还提供了洞察力 转化为细胞可诱导的增强子调控机制。

Deregulation of genome interactions through mutations at regulatory elements or mutation of responsible cellular nuclear enzymes result in multiple disorders. Although the significance of proper genomic-genomic interactions has been seen from disease phenotypes, the underlying molecular mechanisms of genomic looping regulation and the direct outcome of transcription are still not well connected. This partly stems from the highly complex nature of cellular promoter regulation, since it is controlled by multiple transcriptional factors and enhancer elements with significant noise within cell populations. Transcriptomics, genomics, and proteomics studies with recombinant Kaposi's sarcoma-associated herpesvirus (KSHV) episomes identified that the host cellular ChAHP (CHD4, ADNP, HP1) protein complex forms a stable protein complex with KSHV latency associated nuclear antigen (LANA) and plays essential roles in maintaining the inducibility of latent viral chromatin. The ChAHP complex is known to restrict cellular enhancer accessibility and regulates cell lineage, although how the protein complex regulates enhancer-promoter interaction remains unknown. Here, we hypothesize that the KSHV latency-lytic switch is a product of enhancer-promoter interactions regulated by the LANA-ChAHP complex. In this application, we will study how the KSHV episome is maintained as an inducible episome and if we can target the protein complex for therapeutic intervention. The molecular action of the LANA/ChAHP protein complex will be studied from three different perspectives; (i) biochemical, (ii) genetic, and (iii) protein complex structure. By taking advantage of defined enhancer-promoter pairs and convenient inducible mini viral chromatin, we will study enhancer-promoter regulations by dissecting ChAHP protein complex function. The proposed studies should not only benefit the understanding of the KSHV latency-lytic switch, but also provide insight into cellular inducible enhancer regulatory mechanisms.