Center for Structural Biology of HIV RNA

HIV RNA结构生物学中心

• 批准号: 10505794
• 负责人: ALICE TELESNITSKY
• 金额: $ 85.45万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-09 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10505794
• 关键词: Adopted; Affect; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biology; Catalytic RNA; Cells; Collaborations; Complex; CpG dinucleotide; Deamination; Epitopes; Genes; Goals; HIV; HIV Genome; HIV-1; Infection; Interferons; Investigation; Lead; Length; Location; Messenger RNA; Modification; Nucleotides; Participant; Proteins; RNA; RNA Binding; RNA Recognition Motif; RNA Sequences; RNA Stability; RNA-Protein Interaction; Reader; Regulation; Research Personnel; Role; Specific qualifier value; Specificity; Structure; TRIM Gene; TRIM25 gene; TRIM5 gene; Tertiary Protein Structure; Untranslated RNA; Viral; Viral Genome; Virus; Virus Replication; Zinc Fingers; base; cofactor; epitranscriptomics; experimental study; gene product; novel; particle; protein complex; protein function; recruit; screening; sensor; structural biology; viral RNA

Project 2 Summary HIV-1 RNA interacts with many host proteins, some of which enhance and some of which inhibit viral replication. This project will focus on protein:RNA complexes involving HIV-1 RNA and host proteins. Because the HIV-1 genome has all the features of a cellular mRNA, the question of whether and how the HIV-1 genome is differentiated from cellular mRNAs, or ‘sensed’ by host cell proteins is an important principle underpinning this project. Distinguishing features of HIV-1 RNAs include RNA sequence, structures, compositional biases and base modifications. These features of the HIV-1 RNA can enable recognition by antiviral proteins or sensors, which feature prominently in the project. The team will continue its existing collaboration on the structure and function of APOBEC3 proteins, and will build on their prior crystallographic studies of APOBEC3H:RNA interactions to determine how APOBEC3H:RNA binding inhibits HIV-1 infection independently of deamination activity. The team will also determine the structure of an APOBEC3H-Vif complex and will use structural approaches to determine how viral RNA is recognized by additional APOBEC3 proteins. Additionally, the team will build on its prior groundbreaking discoveries of zinc finger antiviral protein (ZAP) and its CpG RNA target sequence to explore the function of protein complexes containing ZAP and its cofactors. They will determine the structures of ZAP domains, the full-length ZAP protein, and complexes containing ZAP with cofactors TRIM25, Riplet and/or KHNYN. They will also use structural biology approaches to determine the mechanistic basis for specific RNA recognition by ZAP, and biological assays to determine the features of optimal CpG-rich ZAP target sequences. The team will also pursue structural, biochemical, and biological investigations of another antiviral protein that binds HIV-1 RNA, TRIM56. The mechanism of action of TRIM56 is poorly understood, thus the team will determine which viral and cellular RNAs are recognized by TRIM56, and how that recognition leads to recruitment of effector functions that affect the fate of targeted RNAs and consequently HIV-1 replication. In addition to detailed studies of known HIV-1 RNA:host protein interactions, the team will also execute a number of exploratory/discovery studies, including various screening approaches to identify cellular proteins that interact with HIV-1 RNA. They will identify novel host proteins that participate in the epitranscriptomic regulation of HIV- 1 RNA fate and investigate the how the m6A reader YTHDF2 regulates RNA stability. The team will also identify RNA-binding interferon-stimulated gene products with anti-HIV-1 activity and zinc finger proteins that affect HIV- 1 replication Finally, the team will explore how the unusually biased HIV-1 RNA nucleotide composition affects viral replication. Thus, the scope of the project encompasses investigations intended to discover novel RNA- protein interactions, as well as studies devoted to atomic level structural characterization of HIV-1 RNA-protein interactions.

项目2总结 HIV-1 RNA 与许多宿主蛋白相互作用，其中一些蛋白增强病毒复制，另一些蛋白抑制病毒复制。 该项目将重点研究涉及 HIV-1 RNA 和宿主蛋白的蛋白质：RNA 复合物。 基因组具有细胞 mRNA 的所有特征，但 HIV-1 基因组是否以及如何存在的问题 与细胞 mRNA 区分开来，或被宿主细胞蛋白“感知”是支撑这一点的重要原则。 HIV-1 RNA 的区分特征包括 RNA 序列、结构、组成偏差和 HIV-1 RNA 的这些特征可以被抗病毒蛋白或传感器识别， 该团队将继续在结构和方面进行现有的合作。 APOBEC3 蛋白的功能，并将建立在他们之前对 APOBEC3H:RNA 的晶体学研究的基础上 相互作用以确定 APOBEC3H:RNA 结合如何独立于脱氨基作用抑制 HIV-1 感染 该团队还将确定 APOBEC3H-Vif 复合物的结构，并使用结构。 此外，该团队还提出了确定病毒 RNA 如何被其他 APOBEC3 蛋白识别的方法。 将建立在其先前对锌指抗病毒蛋白 (ZAP) 及其 CpG RNA 靶标的突破性发现的基础上 序列来探索含有 ZAP 及其辅因子的蛋白质复合物的功能。 ZAP 结构域、全长 ZAP 蛋白以及含有 ZAP 与辅因子 TRIM25 的复合物， Riplet 和/或 KHNYN 还将使用结构生物学方法来确定其机制基础。 ZAP 的特异性 RNA 识别以及生物测定以确定富含 CpG 的最佳 ZAP 靶标的特征 该团队还将对另一种抗病毒药物进行结构、生化和生物学研究。 结合 HIV-1 RNA 的蛋白质 TRIM56 对 TRIM56 的作用机制知之甚少，因此研究小组。 将确定 TRIM56 识别哪些病毒和细胞 RNA，以及这种识别如何导致 效应子功能的募集会影响目标 RNA 的命运，从而影响 HIV-1 的复制。 除了对已知的 HIV-1 RNA：宿主蛋白相互作用的详细研究外，该团队还将执行一些 探索性/发现性研究，包括识别相互作用的细胞蛋白的各种筛选方法 他们将鉴定出参与 HIV-1 表观转录调控的新型宿主蛋白。 1 RNA 命运并研究 m6A 阅读器 YTHDF2 如何调节 RNA 稳定性 该团队还将确定。 RNA结合干扰素刺激的基因产物具有抗HIV-1活性和影响HIV-的锌指蛋白 1 复制 最后，研究小组将探索异常偏向的 HIV-1 RNA 核苷酸组成如何影响 因此，该项目的范围包括旨在发现新型 RNA 的研究。 蛋白质相互作用，以及致力于 HIV-1 RNA 蛋白质原子水平结构表征的研究 互动。