Center for Structural Biology of HIV RNA

项目2摘要 HIV-1RNA与许多宿主蛋白相互作用，其中一些可以增强病毒复制，另一些可以抑制病毒复制。 这个项目的重点是蛋白质：包括HIV-1RNA和宿主蛋白质的RNA复合体。因为HIV-1 基因组具有细胞信使核糖核酸的所有特征，即艾滋病毒-1基因组是否以及如何 区别于细胞内的mRNAs，或被宿主细胞蛋白“感知”，是支持这一点的一个重要原则。 项目。HIV-1 RNA的区别特征包括RNA序列、结构、组成偏见和 碱基修改。HIV-1RNA的这些特征可以使抗病毒蛋白或传感器识别， 它们在项目中占据了突出的地位。该小组将继续其现有的合作结构和 APOBEC3蛋白的功能，并将建立在他们先前的APOBEC3H：RNA的结晶学研究的基础上 相互作用确定APOBEC3H：RNA结合是如何独立于脱氨基抑制HIV-1感染的 活动。该团队还将确定APOBEC3H-Vif复合体的结构，并将使用结构 确定病毒RNA如何被额外的APOBEC3蛋白识别的方法。此外，该团队 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 测序以探索含有ZAP及其辅因子的蛋白质复合体的功能。他们将决定 ZAP结构域、全长ZAP蛋白以及含有ZAP与辅因子TRIM25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 用ZAP识别特异性RNA，并用生物学方法确定最佳富含CpG的ZAP靶标的特征 序列。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 与HIV-1RNA结合的蛋白质，TRIM56。人们对TRIM56的作用机制知之甚少，因此该团队 将决定TRIM56识别哪些病毒和细胞RNA，以及这种识别如何导致 影响靶RNA命运从而影响HIV-1复制的效应器功能的招募。在……里面 除了对已知的HIV-1RNA：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/发现研究，包括识别相互作用的细胞蛋白质的各种筛选方法 携带HIV-1核糖核酸。他们将识别参与HIV表位转录调控的新宿主蛋白- 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 具有抗HIV-1活性的RNA结合干扰素刺激基因产物和影响HIV的锌指蛋白- 最后，研究小组将探索异常偏向的HIV-1RNA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的RNA- 蛋白质相互作用，以及致力于HIV-1RNA-蛋白质原子级结构特征的研究 互动。

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.