RNA post-transcriptional modifications as possible communication hubs between substances of abuse and HIV-1 replication processes

药物滥用是艾滋病毒/艾滋病治疗中的一个恶化因素， 更高的病毒载量、更快的疾病进展和增强的传播。了解 这些增强效应在分子水平上的机制可能导致细胞内的 目前的抗逆转录病毒疗法的管理和新的治疗目标的发现。最近 关于N6-甲基腺苷（m6 A）在HIV-1复制中的活性及其在可卡因中的积累的研究结果- 处理的细胞表明，RNA转录后修饰（PTM）可能是一个关键组成部分， 药物和RNA过程之间广泛的通讯。除了m6 a， 基于质谱（MS）的综合分析方法使我们能够识别 从感染的细胞和病毒体分离的病毒RNA上的许多其他PTM。他们惊人的多样性和 大量的证据支持该提案的假设，即RNA PTM可能介导病毒-宿主通信 通过调节必要的分子相互作用，这可能受到细胞内神经调节剂的影响， 发信号。我们高度创新的方法将使我们能够通过识别RNA来验证这一假设 直接受PTM存在影响的过程，决定多巴胺的作用， GABA信号转导对PTM表达的影响，并最终识别各自之间的重叠 途径，这将揭示细胞内信号传导和病毒之间可能的串扰机制， 感染为此，我们将进行PTM酶和关键调控基因的沉默， 他们的控制网络，同时监测PTM景观和基本指标， 病毒和细胞适应性我们将确定PTM景观的变化在不存在和存在 多巴胺和GABA，同时监测对突出的RNA加工的影响， 基础设施演进然后，我们将探测在两个时间段内显示最显著变化的PTM。 感染和神经调节剂治疗。HIV-1基因组不编码任何已知的 生物遗传酶意味着宿主酶必须负责检测到的病毒PTM，因此 将它们置于关键的细胞内信号的控制之下。修改可以表示对 宿主的代谢基础设施，这使得病毒RNA能够逃避监测和消除的 宿主的防御因此，可以设想，这种假定的适应机制可能是 受到神经调节信号通路的影响。该项目将寻找证据，并将建立 这是指导深入机制调查所必需的框架。改革的影响 拟议的活动将得到全面检验这一假设的能力的证实， 揭示PTM对主要RNA过程的影响，确定其可能的调控机制， 机制，并了解其生物遗传酶在病毒生命周期和适应性中的重要性。

Drug abuse represents an aggravating factor in the treatment of HIV/AIDS, which is associated with higher viral load, faster disease progression, and enhanced transmission. Understanding the mechanisms of these potentiating effects at the molecular level could lead to significant changes in the administration of current anti-retroviral treatments and the discovery of new therapeutic targets. Recent findings on the activity of N6-methyladenosine (m6A) in HIV-1 replication and its accumulation in cocaine- treated cells suggest that RNA post-transcriptional modification (PTM) may represent a key component of extensive communications between drug and RNA processes. In addition to m6A, the implementation of a comprehensive analytical approach based on mass spectrometry (MS) allowed us to identify numerous other PTMs on viral RNA isolated from infected cells and virions. Their striking diversity and abundance support the proposal’s hypothesis that RNA PTMs may mediate virus-host communications by modulating essential molecular interactions, which can be influenced by neuromodulators intracellular signaling. Our highly innovative approach will allow us to test this hypothesis by identifying RNA processes that are directly affected by the presence of PTMs, determining the effects of dopamine and GABA signal transduction on PTM expression, and finally recognizing overlaps between the respective pathways, which will reveal possible crosstalk mechanisms between intracellular signaling and viral infection. To this effect, we will perform silencing of PTM enzymes and key regulatory genes to retrace their control networks, while simultaneously monitoring the PTM landscape and essential indicators of virus and cell fitness. We will determine the variations of PTM landscapes in the absence and presence of dopamine and GABA, while simultaneously monitoring the effects on prominent RNA processing infrastructure. We will then probe the PTMs that displayed the most significant variations during both infection and neuromodulator treatment. The fact that the HIV-1 genome does not code for any known biogenetic enzyme implies that host enzymes must be responsible for the detected viral PTMs, thus placing them under the control of key intracellular signaling. Modification may represent adaptation to the host’s metabolic infrastructure, which enables the viral RNA to evade surveillance and elimination by the host’s defenses. Therefore, it is possible to envision that this putative adaptation mechanism could be affected by neuromodulator signaling pathways. This project will look for evidence and will establish the framework necessary to guide in-depth mechanistic investigations. The transformative impact of the proposed activities will be substantiated by the ability to test this hypothesis in comprehensive fashion, expose the influence of PTMs on prominent RNA processes, identify their possible regulatory mechanisms, and understand the significance of their biogenetic enzymes in viral lifecycle and fitness.