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加重因素，这与 病毒载量更高，疾病进展更快，传播更快。了解 这些增强效应在分子水平上的机制可能导致细胞内 管理目前的抗逆转录病毒治疗和发现新的治疗靶点。近期 HIV-1复制中N6-甲基腺苷(M6A)活性及其在可卡因中蓄积的研究进展 处理过的细胞表明，RNA转录后修饰(PTM)可能是一个关键成分 药物和核糖核酸过程之间的广泛交流。除了m6A之外，实现 一种基于质谱学(MS)的综合分析方法使我们能够识别 从受感染的细胞和病毒粒子中分离的病毒RNA上的许多其他PTM。它们惊人的多样性和 丰度支持了该提议的假设，即RNAPTMS可能介导病毒与宿主之间的通信 通过调节基本的分子相互作用，这可能会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来检验这一假设 直接受PTMS存在影响的过程，确定多巴胺和 GABA信号转导对PTM的表达，最终识别出各自之间的重叠 途径，这将揭示细胞内信号和病毒之间可能的串扰机制 感染。为此，我们将对PTM酶和关键调控基因进行沉默以进行追溯 他们的控制网络，同时监测PTM情况和基本指标 病毒和细胞适合性。我们将确定PTM景观在没有和存在的情况下的变化 多巴胺和GABA，同时监测对显著的RNA加工的影响 基础设施。然后我们将探索在这两个过程中显示最显著变化的PTM 感染和神经调节剂治疗。HIV-1基因组并不编码任何已知的 生物遗传酶意味着宿主酶必须对检测到的病毒PTM负责，因此 将它们置于关键的细胞内信号的控制之下。修改可能代表对 宿主的新陈代谢基础设施，使病毒RNA能够逃避监视和被 主办方的防御。因此，可以设想，这一假定的适应机制可能是 受神经调节剂信号通路的影响。该项目将寻找证据，并将建立 指导深入机械性调查所需的框架。经济衰退带来的变革影响 拟议的活动将通过以全面的方式检验这一假设的能力来证实， 揭示PTMS对重要的RNA过程的影响，确定其可能的调节 机制，并了解它们的生物生成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.