Zika virus nonstructural protein 5 inhibition of interferon signaling

寨卡病毒非结构蛋白 5 对干扰素信号传导的抑制

基本信息

批准号：
10641222
负责人：
Matthew J Evans
金额：
$ 85.42万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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：
2023-08-21 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10641222
关键词：
Amino Acids Animal Model Animals Antiviral Response Antiviral Therapy Attenuated Attenuated Vaccines Binding CRISPR screen Cell Communication Cell Culture Techniques Complex Conserved Sequence Dengue Virus Disease Disease Outbreaks Engineering Evolution Flavivirus Flavivirus Infections Future Genes Genetic Genetic Determinism Goals Human Immune Immune response Immunosuppression In Vitro Infection Innate Immune Response Integration Host Factors Interferon Type I Interferons Knock-out Libraries Maps Mediating Methods Modification Molecular Molecular Virology Mus Mutagenesis Mutate Mutation Nonstructural Protein Orthologous Gene Pathogenesis Pattern Predisposition Process Property Proteins Proteomics Public Health Reporting Resistance Resolution Signal Induction Signal Transduction Site Small Interfering RNA Stat2 protein Structure Surveys Testing Time Tropism Ubiquitin Ubiquitination Vaccine Design Vaccines Variant Viral Viral Genes Virus Work Yellow fever virus Zika Virus antagonist antiviral drug development combat design future outbreak human pathogen in vivo inhibitor innovation insight interferon antagonist mutant mutation screening novel pressure programs recruit response targeted treatment therapeutic target therapy development tool transcription factor ubiquitin ligase vector-borne whole genome zika fever

项目摘要

PROJECT SUMMARY All vector-borne flavivirus NS5 proteins suppress host type I interferon (IFN) signaling, which is critical to successfully infecting humans and causing disease. Inhibitors of this function of NS5 or viruses engineered to lack this activity may be effective antiviral therapies and attenuated vaccines, respectively. However, a deeper understanding of how the flavivirus NS5 protein acts to suppress IFN is needed. Despite the importance of inhibiting IFN signaling, and the high degree of sequence conservation between NS5 proteins within this genus, flaviviruses have evolved numerous distinct ways to antagonize this innate immune response. Many flavivirus NS5 proteins, including those of dengue (DENV), Zika (ZIKV), and yellow fever (YFV) viruses, inhibit the Signal Transducer And Activator Of Transcription 2 (STAT2) protein, which is a transcription factor that mediates IFN signaling. In this application, we propose to fill critical gaps in our understanding of the ZIKV NS5-STAT2 interface by defining the essential viral and host genetic determinants for this interaction at the highest possible, single amino acid, resolution through deep mutational scanning (DMS). In Aim 1, we will screen libraries of all possible single amino acid ZIKV NS5 variants for the ability to suppress IFN signaling. We will examine these determinants in a range of flaviviruses to understand functional conservation and mechanisms. In Aim 2, we will identify how STAT2 genetics impact the ability of ZIKV NS5 to suppress IFN signaling and mediate infection. Finally, in Aim 3 we will define how ZIKV NS5 induces the degradation of human STAT2 by identifying the STAT2 sites where NS5 induces ubiquitination and the host factors ZIKV NS5 recruits to mediate this modification. The results of this project will provide in-depth insights into flavivirus host cell interactions and replication mechanisms that may aid in the development of therapies for ZIKV, and could perhaps be further applied in combating other future virus outbreaks.

项目摘要所有矢量传播黄病毒NS5蛋白抑制宿主I型干扰素（IFN）信号，这对成功感染人类并引起疾病。 NS5的这种功能的抑制剂或设计为缺乏这种活性可能是有效的抗病毒疗法和疫苗减弱的。但是，更深了了解黄病毒NS5蛋白如何作用以抑制IFN。尽管很重要抑制IFN信号传导，以及该属内NS5蛋白之间的高度序列保守，黄病毒已经发展了许多不同的方法来拮抗这种先天免疫反应。许多黄病毒 NS5蛋白，包括登革热（DENV），Zika（Zikv）和黄热病（YFV）病毒的蛋白质，抑制信号转录2（STAT2）蛋白的换能器和激活剂，这是介导IFN的转录因子信号。在此应用程序中，我们建议在对Zikv NS5-Stat2的理解中填补关键空白通过在最高可能的情况下定义这种相互作用的必需病毒和宿主遗传决定因素的界面单氨基酸，通过深突变扫描（DMS）解决。在AIM 1中，我们将筛选所有的库可能的单个氨基酸ZIKV NS5变体，可抑制IFN信号传导的能力。我们将检查这些一系列黄病毒中的决定因素，以了解功能保护和机制。在AIM 2中，我们将确定STAT2遗传学如何影响ZIKV NS5抑制IFN信号传导并介导感染的能力。最后，在AIM 3中，我们将定义ZIKV NS5如何通过识别STAT2来诱导人类Stat2的降解 NS5诱导泛素化和宿主因素ZIKV NS5新兵介导这种修饰的地点。这该项目的结果将提供对黄病毒宿主细胞相互作用和复制机制的深入见解这可能有助于开发ZIKV疗法，也许可以进一步应用未来病毒爆发。