Center for Structural Biology of HIV RNA

HIV RNA结构生物学中心

• 批准号: 10641987
• 负责人: ALICE TELESNITSKY
• 金额: $ 88.72万
• 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/10641987
• 关键词: Adopted; Affect; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biology; Cells; Collaborations; Complex; CpG dinucleotide; Crystallography; Cytoplasm; 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; Viral Physiology; Viral Proteins; 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摘要 HIV-1 RNA与许多宿主蛋白相互作用，其中一些增强病毒复制，一些抑制病毒复制。 该项目将侧重于涉及HIV-1 RNA和宿主蛋白质的蛋白质：RNA复合物。因为HIV-1 基因组具有细胞mRNA的所有特征，HIV-1基因组是否以及如何被 从细胞mRNA中分化出来，或被宿主细胞蛋白质“感知”，是支持这一点的重要原则。 项目HIV-1 RNA的区别特征包括RNA序列、结构、组成偏差和 基地改造HIV-1 RNA的这些特征能够被抗病毒蛋白或传感器识别， 这在项目中非常突出。该小组将继续其现有的结构合作， APOBEC 3蛋白的功能，并将建立在他们以前的APOBEC 3 H：RNA晶体学研究的基础上 确定APOBEC 3 H：RNA结合如何独立于脱氨基作用抑制HIV-1感染的相互作用 活动该团队还将确定APOBEC 3 H-Vif复合物的结构，并将使用结构分析。 确定病毒RNA如何被其他APOBEC 3蛋白识别的方法。此外，该团队 将建立在其先前的锌指抗病毒蛋白（ZAP）及其CpG RNA靶点的突破性发现的基础上 序列，以探索含有ZAP及其辅因子的蛋白质复合物的功能。他们将决定 ZAP结构域、全长ZAP蛋白和含有ZAP与辅因子TRIM 25的复合物的结构， Riplet和/或KHNYN他们还将使用结构生物学方法来确定 ZAP的特异性RNA识别，以及确定最佳富含CpG的ZAP靶标特征的生物测定 序列的该小组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TRIM 56。TRIM 56的作用机制知之甚少，因此研究小组 将确定哪些病毒和细胞RNA被TRIM 56识别，以及这种识别如何导致 募集效应子功能，影响靶向RNA的命运并因此影响HIV-1复制。在 除了对已知的HIV-1 RNA：宿主蛋白质相互作用进行详细研究外，研究小组还将进行一些 探索性/发现性研究，包括各种筛选方法，以确定相互作用的细胞蛋白质 HIV-1 RNA他们将鉴定出参与HIV表观转录调控的新型宿主蛋白， 1 RNA命运，并研究m6 A阅读器YTHDF 2如何调节RNA稳定性。该小组还将确定 RNA结合干扰素刺激的具有抗HIV-1活性的基因产物和影响HIV-1的锌指蛋白 最后，研究小组将探索异常偏置的HIV-1 RNA核苷酸组成如何影响 病毒复制因此，该项目的范围包括旨在发现新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 交互.