Restriction of KSHV by cellular RNA decay pathways

细胞 RNA 衰变途径对 KSHV 的限制

基本信息

批准号：
10699800
负责人：
John Karijolich
金额：
$ 40.03万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10699800
关键词：
3&apos Untranslated Regions Acquired Immunodeficiency Syndrome Address B-Lymphocytes Biochemical Biogenesis Biology Cell model Cells DNA Virus Infections DNA Viruses Data Defense Mechanisms Development Disease Disease Progression Endoribonucleases Ensure Etiology Gammaherpesvirinae Gene Expression Genes Genetic Transcription Herpesviridae Infections Human Human Herpesvirus 8 Immune response Kaposi Sarcoma Knowledge Life Cycle Stages Link Literature Lymphoma cell Lymphoproliferative Disorders Malignant Neoplasms Mass Spectrum Analysis Mediating Messenger RNA Methodology Molecular Outcome Pathway interactions Plants Positioning Attribute Predisposition Process Proteins Quality Control RNA RNA Decay RNA Degradation RNA Processing RNA Viruses Regulation Reporting Retroviridae Ribosomes Role Series Testing Therapeutic Transcript Transcriptional Activation Translations Viral Viral Genes Viral Genome Viral Proteins Virus Virus Replication antagonist antiviral immunity experimental study genome wide screen genome-wide in vivo infected B cell insight lytic replication mRNA Decay member novel therapeutic intervention pathogen primary effusion lymphoma protein complex recombinant virus recruit response transcription factor transcriptome transcriptomics viral RNA

项目摘要

Project Summary Kaposi’s sarcoma-associated herpesvirus (KSHV) is a member of the subfamily gammaherpesvirinae and is causally associated with the development of several malignancies including Kaposi’s sarcoma and primary effusion lymphoma (PEL). The virus’ ability to establish latency as well as reactivate are essential for the development of KSHV-associated disease. Despite these requirements for disease progression there are significant gaps in knowledge regarding cell-intrinsic mechanisms that restrict the KSHV lifecycle. While innate restriction is typically thought of in the context of antiviral immune responses, growing evidence suggests that cellular RNA quality control pathways have an antiviral role. Nonsense-mediated RNA decay (NMD) is an evolutionarily conserved RNA decay pathway that facilitates degradation of RNAs on which ribosomes are deemed to terminate translation aberrantly. Emerging evidence has pointed to a role for NMD in antiviral restriction with a prominent role limiting replication of positive-stranded RNA viruses, however, a role for NMD in DNA virus restriction, such as KSHV, was unknown. We recently reported the discovery that NMD is a cell- intrinsic restriction mechanism for DNA viruses and demonstrated that it imposes a significant restriction on the KSHV lifecycle in PEL cells. Our data demonstrate that NMD-dependent restriction is linked both to the regulation of the unfolded protein response (UPR) as well as targeted degradation of the main KSHV transcription factor, RTA. Building upon these observations our central hypothesis is that NMD restricts KSHV reactivation by targeting key viral mRNAs as well as cellular transcripts in pathways important for viral gene expression and that the virus antagonizes NMD through viral-encoded mechanisms. To test this hypothesis, we propose an integrated series of experiments aimed at determining the interactions between NMD and KSHV. In Aim 1, we will investigate how NMD-dependent regulation of the UPR pathway regulates the KSHV lifecycle. In Aim 2, we will investigate a class of KSHV mRNAs that escape NMD despite harboring sequence features that should render them NMD-susceptible. In Aim 3, we will determine the mechanism by which KSHV-encoded proteins inhibit NMD. Completion of these studies is expected to determine how NMD restricts the KSHV lifecycle as well as the mechanisms the virus employs to antagonize it. These will represent fundamental new insights into how DNA virus infection is regulated by cell-intrinsic mechanisms and can be harnessed for the development of new therapeutic strategies.

项目概要卡波西肉瘤相关疱疹病毒 (KSHV) 是伽玛疱疹病毒亚科的成员，是与卡波西肉瘤和原发性肉瘤等多种恶性肿瘤的发生有因果关系病毒建立潜伏期和重新激活的能力对于渗出性淋巴瘤（PEL）至关重要。尽管有这些对疾病进展的要求，但仍存在 KSHV 相关疾病的发展。关于限制 KSHV 生命周期的细胞内在机制的知识存在重大差距。限制通常被认为是在抗病毒免疫反应的背景下，越来越多的证据表明细胞 RNA 质量控制途径具有抗病毒作用，无义介导的 RNA 衰减 (NMD) 是一种。进化上保守的 RNA 降解途径，促进核糖体所在 RNA 的降解被认为异常终止翻译的新证据表明 NMD 在抗病毒方面的作用。限制性限制具有限制正链RNA病毒复制的突出作用，然而，NMD在 DNA 病毒限制，例如 KSHV，是未知的。我们最近报道了 NMD 是一种细胞-病毒的发现。 DNA病毒的内在限制机制，并证明它对DNA病毒施加了显着的限制 PEL 细胞中的 KSHV 生命周期表明 NMD 依赖性限制与调节有关。未折叠蛋白反应 (UPR) 以及主要 KSHV 转录因子的定向降解， RTA。基于这些观察，我们的中心假设是 NMD 通过以下方式限制 KSHV 重新激活：针对病毒基因表达重要途径中的关键病毒 mRNA 以及细胞转录本病毒通过病毒编码机制对抗 NMD。为了检验这一假设，我们提出了一个假设。在目标 1 中，我们进行了旨在确定 NMD 和 KSHV 之间相互作用的一系列综合实验。在目标 2 中，我们将研究 UPR 途径的 NMD 依赖性调节如何调节 KSHV 生命周期。将研究一类逃脱 NMD 的 KSHV mRNA，尽管其序列特征应该使它们对 NMD 敏感。在目标 3 中，我们将确定 KSHV 编码蛋白的机制。抑制 NMD 预计将确定 NMD 如何限制 KSHV 生命周期。作为病毒对抗病毒的机制，这些将代表关于如何对抗它的基本新见解。 DNA 病毒感染受细胞内在机制调节，可用于开发新的治疗策略。