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

• 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
• 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

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.

项目总结 所有载体携带的黄病毒NS5蛋白都抑制宿主I型干扰素(IFN)信号，这对 成功感染人类并引发疾病。NS5这一功能的抑制剂或经工程改造的病毒 缺乏这种活性可能分别是有效的抗病毒疗法和减毒疫苗。然而，更深一层的 了解黄病毒NS5蛋白如何抑制干扰素是很有必要的。尽管重要的是 抑制干扰素信号，以及该属内NS5蛋白之间的高度序列保守性， 黄病毒已经进化出许多不同的方法来对抗这种先天免疫反应。许多黄病毒 NS5蛋白，包括登革热(DENV)、寨卡病毒(ZIKV)和黄热病(YFV)病毒的蛋白，抑制信号 转录转导和激活因子2(STAT2)蛋白，它是一种介导干扰素的转录因子 发信号。在本申请中，我们建议填补我们对ZIKV NS5-STAT2的理解中的关键空白 通过尽可能高地定义这种相互作用的基本病毒和宿主遗传决定因素， 单一氨基酸，通过深度突变扫描(DMS)进行分辨。在目标1中，我们将筛选所有库 可能的单一氨基酸ZIKV NS5变种具有抑制干扰素信号的能力。我们将对这些进行研究 了解一系列黄病毒的决定因素，以了解功能保守和机制。在目标2中，我们将 确定STAT2基因如何影响ZIKV NS5抑制干扰素信号和介导感染的能力。 最后，在目标3中，我们将定义ZIKV NS5如何通过鉴定STAT2来诱导人类STAT2的降解 NS5诱导泛素化的部位和宿主因子ZIKV NS5招募新成员来调节这种修饰。这个 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了解 这可能有助于开发ZIKV的治疗方法，并可能进一步应用于对抗其他 未来的病毒暴发。