Mechanisms mediating immune response upon sensing of nuclear viral DNA

感测核病毒 DNA 介导免疫反应的机制

基本信息

批准号：
10672215
负责人：
Ileana M. Cristea
金额：
$ 32.38万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10672215
关键词：
Adaptor Signaling Protein Address Antiviral Response Belief Binding Biochemical Biological Assay Biology Biophysics Cell Nucleus Cells Chromatin Complex Cytomegalovirus DNA DNA Binding DNA Damage DNA Repair DNA Virus Infections DNA Viruses DNA biosynthesis DNA-dependent protein kinase Event Funding Gene Expression Gene Expression Regulation Genetic Transcription Genome Health Herpesviridae Herpesviridae Infections Herpesvirus 1 Heterochromatin Holoenzymes Host Defense Human Immune Immune Evasion Immune System Diseases Immune response Immune signaling Immune system Immunity Immunosuppression Infection Innate Immune Response Innate Immune System Interferons Knowledge LOXL2 gene Link Liquid substance Location Lytic Phase Mammalian Cell Mass Spectrum Analysis Mediating Modeling Natural Immunity Nonhomologous DNA End Joining Nuclear Pathway interactions Phase Phase Transition Phosphorylation Phosphotransferases Physical condensation Pilot Projects Polymerase Process Promoter Regions Property Proteins Proteome RNA Polymerase II Regulation Reporting Repression Research Shapes Signal Transduction Specificity Testing Transcription Regulation Pathway Viral Viral Genes Viral Genome Virus Virus Diseases Virus Replication carcinogenesis combat cytokine genetic approach human DNA human pathogen live cell microscopy marenostrin optogenetics pathogen pathogenic virus receptor recruit response sensor transmission process viral DNA

项目摘要

In order to mount intrinsic and innate immune responses to infections by DNA viruses, mammalian cells rely on specialized proteins that recognize the viral DNA as a foreign molecule. Upon binding to viral DNA, these sensors induce cytokine secretion, prompting neighboring cells to activate their defenses and inhibiting the spread of infection. Recent years have seen significant progress in the understanding of processes governing viral DNA sensing. Contrary to prior dogma, we determined that mammalian cells can distinguish viral DNA from self-DNA in the nuclei of infected cells. The characterization of the interferon inducible protein IFI16 as the first known nuclear sensor of viral DNA has opened a new research direction in immunity, starting to shed light on how cells detect nuclear-replicating viruses, such as herpesviruses. We further uncovered that the human pathogens, herpes simplex virus type 1 (HSV-1) and human cytomegalovirus (HCMV) have immune evasion mechanisms that specifically suppress IFI16. With this knowledge, during the previously funded R01, we aimed to dissect the mechanisms underlying nuclear IFI16 DNA sensing. We addressed questions regarding where and when the IFI16-viral DNA binding event occurs, the properties that allow IFI16 to sense DNA, and the functional interactions that support IFI16 antiviral responses. Our results uncovered that nuclear DNA sensing relies on dynamic on/off sensor associations with parental viral DNA at the nuclear periphery. We demonstrated that IFI16 oligomerization on viral DNA is essential for nucleus-derived immune signaling. We further established that, upon binding to viral DNA, IFI16 triggers both cytokine expression and suppression of viral gene expression. Our proposal will address several fundamental questions regarding nuclear DNA sensing that have emerged from these findings. In Aim 1, we will define what mechanisms drive dynamic IFI16 oligomerization with HSV-1 DNA at the nuclear periphery. We will test our hypothesis that this property is biophysically conferred via rapid and reversible liquid-phase condensation events, and delineate how these events govern innate immunity and viral replication. Next, we will characterize mechanisms underlying the two antiviral IFI16 functions downstream of its binding to DNA. In Aim 2, we will determine how IFI16 induces innate immune signals upon nuclear DNA sensing. We discovered that IFI16 interacts with and activates the DNA dependent protein kinase (DNA-PK) holoenzyme in response to herpesvirus infections. We will define how IFI16 and the DNA damage response coordinate to stimulate innate immunity and antiviral non- homologous end-joining. In Aim 3, we will elucidate the mechanisms underlying IFI16 restriction of virus gene expression. We will functionally characterize the IFI16 interactions with chromatin modulators that we showed to act as HSV-1 restriction factors. We will systematically define the pathways linking IFI16 to viral genome heterochromatinization. Collectively, our results will characterize a newly discovered aspect of biology that links innate immunity to nuclear processes governing DNA damage repair and gene expression regulation.

为了使DNA病毒对感染的内在和先天免疫反应安装，哺乳动物细胞依靠识别病毒DNA是外国分子的专门蛋白质。与病毒DNA结合后，这些传感器诱导细胞因子分泌，促使相邻细胞激活其防御能力并抑制感染传播。近年来在理解管理过程中取得了重大进展病毒DNA感应。与先前的教条相反，我们确定哺乳动物细胞可以区分病毒DNA 从被感染细胞的核中的自动-DNA。干扰素诱导蛋白IFI16的表征为病毒DNA的第一个已知的核传感器已开辟了免疫力的新研究方向，开始发光关于细胞如何检测核重复病毒，例如疱疹病毒。我们进一步发现了人类病原体，单纯疱疹病毒1型（HSV-1）和人类巨细胞病毒（HCMV）具有免疫逃避专门抑制IFI16的机制。有了这些知识，在先前资助的R01期间，我们的目标剖析核IFI16 DNA感应的基础机制。我们解决了有关哪里的问题当发生IFI16病毒DNA结合事件时，允许IFI16感知DNA的特性，并且支持IFI16抗病毒反应的功能相互作用。我们的结果发现了核DNA感应依赖于核周围的父母病毒DNA的动态开/关。我们证明病毒DNA上的IFI16寡聚化对于核衍生的免疫信号传导至关重要。我们进一步确定，在与病毒DNA结合后，IFI16触发了细胞因子的表达和抑制病毒基因表达。我们的建议将解决有关核DNA的几个基本问题从这些发现中得出的感觉。在AIM 1中，我们将定义哪些机制驱动动态IFI16 用核外围的HSV-1 DNA寡聚。我们将检验我们的假设，即该属性是通过快速和可逆的液相凝结事件赋予生物物理，并描述这些方式事件控制先天免疫和病毒复制。接下来，我们将表征这两个机制抗病毒IFI16在其与DNA结合的下游功能。在AIM 2中，我们将确定IFI16如何诱导核DNA感应时先天免疫信号。我们发现IFI16与并激活 DNA依赖性蛋白激酶（DNA-PK）全酶响应疱疹病毒感染。我们将定义 IFI16和DNA损伤反应如何刺激先天免疫和抗病毒非 - 同源最终连接。在AIM 3中，我们将阐明IFI16病毒基因限制的机制表达。我们将在功能上表征我们显示的IFI16与染色质调节剂的相互作用充当HSV-1限制因素。我们将系统地定义将IFI16与病毒基因组联系起来的途径异染色性。总的来说，我们的结果将描述一个新发现的生物学方面将先天免疫与管理DNA损伤修复和基因表达调控的核过程联系起来。

项目成果

期刊论文数量（63）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Intercellular communication within the virus microenvironment affects the susceptibility of cells to secondary viral infections.

DOI：
10.1126/sciadv.adg3433
发表时间：
2023-05-10
期刊：
SCIENCE ADVANCES
影响因子：
13.6
作者：
Song, Bokai;Sheng, Xinlei;Justice, Joshua L.;Lum, Krystal K.;Metzger, Peter J.;Cook, Katelyn C.;Kostas, James C.;Cristea, Ileana M.
通讯作者：
Cristea, Ileana M.

Protein lipoylation: an evolutionarily conserved metabolic regulator of health and disease.

DOI：
10.1016/j.cbpa.2017.11.003
发表时间：
2018-03
期刊：
Current opinion in chemical biology
影响因子：
7.8
作者：
Rowland EA;Snowden CK;Cristea IM
通讯作者：
Cristea IM

The DNA Sensor IFIX Drives Proteome Alterations To Mobilize Nuclear and Cytoplasmic Antiviral Responses, with Its Acetylation Acting as a Localization Toggle.

DOI：
10.1128/msystems.00397-21
发表时间：
2021-06-29
期刊：
mSystems
影响因子：
6.4
作者：
Howard TR;Crow MS;Greco TM;Lum KK;Li T;Cristea IM
通讯作者：
Cristea IM

Infection-Induced Peroxisome Biogenesis Is a Metabolic Strategy for Herpesvirus Replication.

DOI：
10.1016/j.chom.2018.09.002
发表时间：
2018-10-10
期刊：
Cell host & microbe
影响因子：
30.3
作者：
Jean Beltran PM;Cook KC;Hashimoto Y;Galitzine C;Murray LA;Vitek O;Cristea IM
通讯作者：
Cristea IM

Lamin B1 acetylation slows the G1 to S cell cycle transition through inhibition of DNA repair.