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.

项目概要 所有载体传播的黄病毒 NS5 蛋白都会抑制宿主 I 型干扰素 (IFN) 信号传导，这对于 成功感染人类并引起疾病。 NS5 或病毒的这种功能的抑制剂 缺乏这种活性的可能分别是有效的抗病毒疗法和减毒疫苗。然而，更深层次的 需要了解黄病毒 NS5 蛋白如何抑制 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性， 黄病毒已经进化出许多不同的方式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)、寨卡 (ZIKV) 和黄热病 (YFV) 病毒的 NS5 蛋白，抑制信号 转录转导子和激活子 2 (STAT2) 蛋白，是介导 IFN 的转录因子 发信号。在此应用中，我们建议填补我们对 ZIKV NS5-STAT2 理解的关键空白 通过尽可能最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) 进行解析。在目标 1 中，我们将筛选所有的库 可能的单氨基酸 ZIKV NS5 变体能够抑制 IFN 信号传导。我们将检查这些 一系列黄病毒的决定因素，以了解功能保护和机制。在目标 2 中，我们将 确定 STAT2 遗传学如何影响 ZIKV NS5 抑制 IFN 信号传导和介导感染的能力。 最后，在目标 3 中，我们将通过识别 STAT2 来定义 ZIKV NS5 如何诱导人类 STAT2 降解 NS5 诱导泛素化的位点以及 ZIKV NS5 招募的宿主因子来介导这种修饰。这 该项目的结果将深入了解黄病毒宿主细胞相互作用和复制机制 这可能有助于开发 ZIKV 疗法，并可能进一步应用于对抗其他疾病 未来病毒爆发。