m6A mRNA reader proteins in the AIDS-opportunistic pathogen Toxoplasma gondii

艾滋病机会致病菌弓形虫中的 m6A mRNA 阅读器蛋白

• 批准号: 10615374
• 负责人: William J Sullivan
• 金额: $ 19.51万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-03 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10615374
• 关键词: Acquired Immunodeficiency Syndrome; Acute; Address; Adenosine; Antiparasitic Agents; Binding; Binding Proteins; Binding Sites; Biology; Cell Nucleus; Chronic Disease; Cleavage And Polyadenylation Specificity Factor; Comprehension; Cyst; Cytosol; Data; Development; Family; Felis catus; Follow-Up Studies; Food Contamination; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; HIV/AIDS; Human; Hybrids; Immune response; Immunity; Immunocompromised Host; Immunoprecipitation; Individual; Infection; Knowledge; Life; Mediating; Messenger RNA; Metabolism; Methylation; Modification; Mutation Analysis; Nuclear; Nuclear Export; Opportunistic Infections; Parasites; Patients; Pharmaceutical Preparations; Phenotype; Plants; Polyadenylation Pathway; Population; Positioning Attribute; Process; Proliferating; Proteins; RNA; RNA Probes; RNA Splicing; Reader; Recurrence; Role; Signal Transduction; System; Tertiary Protein Structure; Time; Tissues; Toxoplasma; Toxoplasma gondii; Toxoplasmosis; Transcript; Translations; Untranslated Regions; Water; Writing; Zinc Fingers; combat; crosslink; effective therapy; epitranscriptomics; insight; knock-down; new therapeutic target; novel; novel therapeutics; opportunistic pathogen; polyadenylated messenger RNA; recruit; therapeutic target; trafficking; transcriptome; transmission process

Toxoplasma gondii is an intracellular parasite that causes life-threatening opportunistic infection in HIV/AIDS patients. The replicative stage (tachyzoite) develops into a latent stage (bradyzoite) that is impervious to immunity and approved antiparasitic drugs. Tissue cysts give rise to recurrent reactivation of infection in the immunocompromised, creating chronic disease in HIV/AIDS patients. Compounding this problem is a paucity of safe and effective therapies, which underscores the urgent need to identify essential processes in the parasite that could be exploited for the development of better drugs. We and others recently discovered that methylation of adenosines at position 6 (m6A) is abundant in Toxoplasma mRNA, representing a new layer of gene regulation called epitranscriptomics. Importantly, the proteins that “write” and “read” m6A modifications are essential for parasite viability. Moreover, as this machinery resembles plants more than humans, it represents an attractive new drug target. To study this vulnerability in Toxoplasma, we aim to fill the gap in our knowledge regarding how this signal dictates the fate of mRNA transcripts through the study of m6A reader proteins. We hypothesize that Toxoplasma m6A mRNA reader proteins coordinate different aspects of mRNA metabolism essential for parasite viability. In other species, m6A readers are found in the nucleus and cytosol, regulating the fate of mRNA by modulating splicing, trafficking, and translation. To date, only two nuclear YTH family m6A readers have been identified and they remain largely uncharacterized. And, despite the abundance of m6A mRNA in the cytosol, no cytosolic m6A readers have been identified. We propose two specific aims that will address our hypothesis by answering these questions. Aim 1 will determine the roles of the two plant-like YTH m6A readers operating in the Toxoplasma nucleus. Aim 2 will identify novel m6A reader proteins from tachyzoites and bradyzoites using a functional m6A-binding probe that we developed. These pioneering studies will mark the first detailed analysis of m6A readers in both replicative and latent stages of Toxoplasma, which promises to reveal new therapeutic options to treat this opportunistic infection of HIV/AIDS patients.

弓形虫是一种细胞内寄生虫，可引起危及生命的机会性感染 HIV/艾滋病患者的复制期（速殖子）发展为潜伏期（缓殖子）。 不受免疫和批准的抗寄生虫药物的影响，会产生组织囊肿。 免疫功能低下者感染反复激活，造成慢性疾病 艾滋病毒/艾滋病患者缺乏安全有效的治疗方法，使这一问题变得更加复杂。 这强调了迫切需要确定寄生虫中可能的基本过程 我们和其他人最近发现，这一点被用于开发更好的药物。 弓形虫 mRNA 中 6 位腺苷 (m6A) 的甲基化丰富，代表 一个新的基因调控层，称为表观转录组学，重要的是“书写”的蛋白质。 此外，“读取”m6A 修饰对于寄生虫的生存能力至关重要。 它比人类更类似于植物，因此它代表了一个有吸引力的新药物靶标。 弓形虫的脆弱性，我们的目标是填补我们关于该信号如何发出的知识空白 通过 m6A 阅读器蛋白的研究决定 mRNA 转录本的命运。 弓形虫 m6A mRNA 阅读器蛋白协调 mRNA 代谢的不同方面 在其他物种中，m6A 阅读器存在于细胞核和细胞核中。 细胞质，通过调节剪接、运输和翻译来调节 mRNA 的命运。 仅确定了两个核心 YTH 家族 m6A 阅读器，并且它们大部分仍然存在 并且，尽管细胞质中存在丰富的 m6A mRNA，但没有细胞质 m6A。 我们提出了两个具体目标来解决我们的假设。 回答这些问题的目标 1 将确定两种类似植物的 YTH m6A 的作用。 在 Aim 2 细胞核中运行的读取器将从蛋白质中识别出新型 m6A 读取器。 使用我们开发的功能性 m6A 结合探针进行速殖子和缓殖子。 开创性研究将标志着 m6A 阅读器在复制和复制方面的首次详细分析 弓形虫的潜伏期，有望揭示治疗这种疾病的新治疗方案 HIV/AIDS 患者的机会性感染。