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Gondii是一种细胞内寄生虫，会引起威胁生命的选择感染 在艾滋病毒/艾滋病患者中。复制阶段（tachyzoite）成长为潜在阶段（bradyzoite） 这对免疫学和批准的抗寄生虫药物无关。组织囊肿会产生 在免疫功能低下的感染复发，从而产生慢性病 艾滋病毒/艾滋病患者。加重此问题的是对安全有效的疗法的匮乏， 这突显了迫切需要识别寄生虫中的基本过程 利用用于开发更好的药物。我们和其他人最近发现 位置6（M6A）处腺苷的甲基化在弓形虫mRNA中很丰富，代表 一个新的基因调节层，称为表面参考组学。重要的是，“写”的蛋白质 “读取” M6A修改对于寄生虫的生存能力至关重要。而且，作为此机械 比人类更像植物，它代表了一个有吸引力的新药靶标。研究这个 毒品中的脆弱性，我们旨在填补有关该信号的知识的空白 通过研究M6A读取器蛋白来决定mRNA转录本的命运。我们假设 弓形虫m6a mRNA读取器蛋白坐标mRNA代谢的不同方面 寄生虫生存能力必不可少。在其他物种中，M6A读取器在细胞核中发现，并且 细胞质，通过调节剪接，运输和翻译来调节mRNA的命运。迄今为止， 仅确定了两个核Yth家庭M6A读者，并且在很大程度上仍然存在 未表征。尽管M6a mRNA在细胞质中抽象，但无细胞质M6A 已经确定了读者。我们提出了两个具体目标，这些目标将通过 回答这些问题。 AIM 1将确定两个类似植物的Yth M6A的作用 在弓形虫核中运作的读者。 AIM 2将从 使用我们开发的功能性M6A结合探针的速氮族和曲二氮化岩。这些 开创性研究将标志着复制和复制和 弓形虫的潜在阶段有望揭示新的治疗选择来治疗这一点 艾滋病毒/艾滋病患者的机会感染。