Regulation and Herpes simplex virus 1 counter-regulation of transcriptional bursting kinetics in the early type I interferon response

单纯疱疹病毒1在早期I型干扰素反应中转录爆发动力学的调节和反调节

• 批准号: 470667831
• 负责人: Professor Dr. Lars Dölken
• 金额: --
• 依托单位: Institut für Virologie und Immunbiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/470667831?language=en
• 关键词: Regulation; Herpes; simplex; virus; counter

Type I interferons (IFN) induce the expression of hundreds of genes that augment control of invading viruses. Efficient evasion thereof is crucial for productive virus infection and long-term persistence. We discovered that the induction of interferon-stimulated genes (ISGs) in the first two hours of cytomegalovirus infection predominantly results from an increased likelihood of transcriptional bursts. The magnitude of bursts, however, was unchanged compared to basal expression of ISGs in uninfected cells. Recently, the DTX3L/PARP9 ubiquitin ligase complex was shown to bind to the STAT1/STAT2/IRF9 (ISGF3) complex and ubiquitinate histones within ISG promoters, thereby enhancing the expression of a large subset of ISGs. Interestingly, we found that the large HSV 1 tegument protein pUL36 targets DTX3L/PARP9 with its N terminal deubiquitinating (DUB) domain and inhibits two DTX3L/PARP9-attributed ubiquitin-mediated functions, namely the induction of ISGs, and the recruitment of p53 binding protein 1 (53BP1) to sites of DNA damage. We hypothesize that (i) DTX3L/PARP9, which is recruited to ISG promoters via the ISGF3 complex, facilitates switching ISG promoters from a non-permissive to a permissive state and thereby induces more frequent transcriptional bursts, and (ii) that the HSV-1 pUL36 DUB counteracts this to augment productive infection. The main objective of this project is to elucidate the mechanistic role of the pUL36 DUB on the DXT3L/PARP9-ISGF3 interaction in governing the gene-specific transcriptional output of ISGs. The obtained data will illuminate an exciting new cellular mechanism by which ISGs are regulated at the level of transcriptional bursting and how HSV-1 interferes with it. Finally, we will employ our scSLAM-Seq approach and develop the required computational framework to study ISG bursting kinetics at single cell level. In summary, we will study viral manipulation of ISG induction to elucidate novel cellular mechanisms that govern ISG bursting kinetics at chromatin level.

I 型干扰素 (IFN) 诱导数百个基因的表达，增强对入侵病毒的控制。有效规避其对于有效的病毒感染和长期持续存在至关重要。我们发现巨细胞病毒感染的前两个小时内干扰素刺激基因（ISG）的诱导主要是由于转录爆发的可能性增加所致。然而，与未感染细胞中 ISG 的基础表达相比，爆发的幅度没有变化。最近，DTX3L/PARP9 泛素连接酶复合物被证明可以与 STAT1/STAT2/IRF9 (ISGF3) 复合物结合，并泛素化 ISG 启动子内的组蛋白，从而增强大部分 ISG 的表达。有趣的是，我们发现大型 HSV 1 被膜蛋白 pUL36 以其 N 末端去泛素化 (DUB) 结构域靶向 DTX3L/PARP9，并抑制两个 DTX3L/PARP9 归因的泛素介导的功能，即 ISG 的诱导和 p53 结合蛋白 1 (53BP1) 募集到 DNA 损伤位点。我们假设 (i) DTX3L/PARP9 通过 ISGF3 复合物被招募到 ISG 启动子，促进 ISG 启动子从非允许状态切换到允许状态，从而诱导更频繁的转录爆发，以及 (ii) HSV-1 pUL36 DUB 抵消这种情况以增强生产性感染。该项目的主要目标是阐明 pUL36 DUB 对 DXT3L/PARP9-ISGF3 相互作用在控制 ISG 基因特异性转录输出中的机制作用。获得的数据将阐明一种令人兴奋的新细胞机制，通过该机制，ISG 在转录爆发水平上受到调节，以及 HSV-1 如何干扰它。最后，我们将采用 scSLAM-Seq 方法并开发所需的计算框架来研究单细胞水平的 ISG 爆发动力学。总之，我们将研究病毒对 ISG 诱导的操纵，以阐明在染色质水平上控制 ISG 爆发动力学的新细胞机制。