Ribosome Heterogeneity in Cryptococcus neoformans

新型隐球菌的核糖体异质性

基本信息

批准号：
10494146
负责人：
John C Panepinto
金额：
$ 38.44万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-24 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10494146
关键词：
5&apos Untranslated Regions Affect Antifungal Agents Antifungal Therapy Binding Biology Catalytic Domain Cellular Stress Cessation of life Complex Cryptococcus Cryptococcus neoformans Data Defect Development Drug Targeting Eukaryota Exhibits Fungal Drug Resistance Future Goals Growth Heterogeneity Homologous Gene Human Hypoxia Immune system Immunologic Deficiency Syndromes Impairment In Vitro Intervention Investigation Lung Messenger RNA Molecular Chaperones Mycoses Names Oxidative Stress Pathogenesis Pathway interactions Patients Peptide Initiation Factors Persons Pharmacologic Substance Phenotype Phosphorylation Process Prokaryotic Cells Protein Biosynthesis Protein Isoforms Proteins Proteomics RNA RNA Recognition Motif RNA-Binding Proteins RNA-Protein Interaction Regulation Regulon Resistance Ribosomes Role Sedimentation process Signal Pathway Specificity Stress Structure Techniques Temperature Testing Therapeutic Translating Translation Initiation Translations Transplantation Virulence Work antimicrobial antimicrobial drug base biological adaptation to stress comorbidity experimental study fungus human pathogen knock-down mRNA Decay mRNA capping mortality mutant novel pathogen pathogenic fungus recruit scaffold stress tolerance translation factor translatome

项目摘要

Invasive fungal infections are a major cause of co-morbidity and mortality in patients living with immunodeficiency and transplantation. A limited antifungal arsenal and emergence of antifungal drug resistance have made the need for novel antifungal therapies paramount. Cellular protein synthesis carried out by the ribosome is a known and well-vetted target for pharmaceutical intervention in prokaryotes and eukaryotes, but has not been exploited in antifungal development because of the presumed inability to target the core catalytic function of the ribosome with specificity in fungi. Using the human pathogen Cryptococcus neoformans, we have leveraged two approaches to identify factors that associate with the fungal ribosome under stress conditions encountered in the host. Using a homology based approach, we have identified Cryptococcus-specific homologues of the conserved eukaryotic mRNA cap-binding complex (eIF4F) that we have names alternative eIF4G (AFG1) and alternative eIF4E (AFE1). Deletion of AFG1 results impairs hypoxia adaptation, which is a major pathogenesis-associated pathway in the major human fungal pathogens. Further work on the function of Afg1 and Afe1 as components of a novel eIF4F complex are outlined in Aim 1, and the identification of the mRNA targets they bind is outlined in Aim 3. Our second approach developed a novel proteomic pipeline that we’ve termed RiboPROT to identify the factors associating with translating ribosomes during adaptation to temperature stress and oxidative stress. Aim 2 of this proposal will investigate the role of these proteins in stress-responsive translation and pathogenesis. We have prioritized hits from our initial RiboPROT experiments to investigate further, and propose to identify additional factors under additional conditions relevant to cryptococcal pathogenesis and biology. For those with RNA-binding domains, cognate mRNA targets will be identified in Aim 3, identifying stress responsive translational regulons. At the conclusion of these studies, we will have a comprehensive set of ribosome-associating factors to investigate in future work as antifungal targets. These unique stress-specific and fungi-specific ribosome-associating factors may provide a novel pathway to targeting the fungal ribosome with specificity.

侵袭性真菌感染是免疫缺陷和移植患者合并症和死亡的主要原因。有限的抗真菌库和抗真菌药物耐药性的出现使得对由核糖体进行的新型抗真菌疗法的需求变得至关重要。原核生物和真核生物药物干预的已知且经过严格审查的目标，但由于推测无法针对核心而尚未在抗真菌开发中得到利用使用人类病原体新型隐球菌，我们利用两种方法来鉴定在宿主遇到的应激条件下与真菌核糖体相关的因子，我们使用基于同源性的方法鉴定了隐球菌。保守的真核 mRNA 帽结合复合物 (eIF4F) 的特定同源物，我们将其命名为替代 eIF4G (AFG1) 和替代 eIF4E (AFE1)。删除 AFG1 结果会损害缺氧适应，这是主要人类真菌病原体的主要发病机制相关途径。目标 1 概述了 Afg1 和 Afe1 作为新型 eIF4F 复合物的功能的进一步研究。目标 3 概述了它们结合的 mRNA 靶点的识别。我们的第二种方法开发了一种新型蛋白质组管道，我们将其称为 RiboPROT，以识别与该提案的目标 2 将研究这些蛋白质在应激反应翻译和发病机制中的作用，以进一步研究我们最初的 RiboPROT 实验中的结果，并建议确定其他因素。在与隐球菌发病机制和生物学相关的其他条件下，对于那些具有 RNA 结合结构域的人，将在目标 3 中鉴定同源 mRNA 靶点，并在结论中鉴定应激反应性翻译调节子。通过这些研究，我们将拥有一套全面的核糖体相关因子，可以在未来的工作中作为抗真菌靶标进行研究。这些独特的应激特异性和真菌特异性核糖体相关因子可能会提供一种特异性靶向真菌核糖体的新途径。