The role of C. neoformans Hog1 and its effectors in translatome reprogramming

新型隐球菌Hog1及其效应子在翻译组重编程中的作用

• 批准号: 10679476
• 负责人: David Goich
• 金额: $ 3.4万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679476
• 关键词: Acute; Advisory Committees; Ascomycota; Attenuated; Biogenesis; Biological Assay; Biotin; Biotinylation; Body Temperature; Buffers; Cessation of life; Communication; Compensation; Complement; Cryptococcus neoformans; Data; Defect; Development; Disease; EIF-2alpha; Educational workshop; Equipment; Fungi Model; Genetic Transcription; Goals; Immune system; Infection; Knock-out; Knowledge; Ligase; MAP Kinase Modules; Macrophage; Maps; Masks; Mediating; Messenger RNA; Modeling; Molecular; Mus; Northern Blotting; Nucleic Acid Binding; Nutrient; Osmosis; Output; Oxidative Stress; Pathogenesis; Pathogenicity; Pathway interactions; Persons; Phenotype; Phosphorylation; Phosphotransferases; Play; Process; Protein Biosynthesis; Proteins; Regulation; Regulatory Pathway; Regulon; Repression; Research Personnel; Research Training; Ribosomal RNA; Ribosomes; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Stress; Temperature; Testing; Training; Transcript; Transcriptional Regulation; Translating; Translation Initiation; Translational Regulation; Translational Repression; Universities; Virulence; Work; biological adaptation to stress; candidate identification; design; experimental study; follow-up; human pathogen; improved; insight; liquid chromatography mass spectrometry; mRNA Stability; mimetics; mouse model; mutant; opportunistic pathogen; p38 Mitogen Activated Protein Kinase; pathogenic fungus; polysome profiling; prevent; response; ribosome profiling; skills; stress tolerance; stressor; symposium; training opportunity; transcription factor; transcriptome; transcriptome sequencing; translation factor; translatome; uptake

Abstract Cryptococcus neoformans is an environmental fungus and opportunistic pathogen of people with compromised immune systems that causes an estimated 181,000 deaths annually. A key step in C. neoformans pathogenesis is adaptation to the host, which is mediated by several signal transduction pathways. Translatome reprogramming, changes in protein synthesis, is a key output of these signal transduction pathways that allows C. neoformans to rapidly fine-tune the stress response, and defects in this process are associated with reduced virulence. The Hog1 p38 MAP kinase (MAPK) module is one signaling module that mediates this process, though the mechanism and effectors by which Hog1 mediates reprogramming are unknown in C. neoformans. We propose to identify the molecular basis of Hog1 activation, and generate a high throughput characterization of its interactome. We also found that another translatome regulatory pathway, the Gcn2 pathway, is differentially regulated in hog1∆, and aim to evaluate the implications of this change in both translatome reprogramming as well as virulence. Thus, we hypothesize that Hog1 mediates changes to a broad interactome via its kinase activity, and that crosstalk with the Gcn2 pathway masks more severe defects in stress tolerance and virulence in hog1∆. We will be testing two specific aims: (1) Determine the mechanism by which Hog1 regulates stress responses in C. neoformans, and (2) Evaluate whether increased eIF2α phosphorylation compensates for loss of Hog1. This project is significant as it will elucidate the mechanism of reprogramming by Hog1, will identify effectors that could be targeted to inhibit this process, and will also examine a convergence point between two major signaling pathways. The overall goal of this research training plan is to equip the candidate with the necessary skills to answer these questions, and to provide them with relevant training to establish themselves as a successful academic investigator studying fungal pathogenesis and stress adaptation. These goals will be accomplished under the guidance of a sponsor and thesis advisory committee, with the help of training opportunities and equipment provided by the University at Buffalo, and through professional development opportunities at conferences and workshops.

抽象的 加密赛车是一种环境真菌和受损的人的机会性病原体 免疫系统每年估计会导致181,000人死亡。 Neoformans发病机理的关键步骤 是对宿主的适应，该宿主是由几个信号转导途径介导的。翻译 重编程，蛋白质合成的变化，是这些信号转导途径的关键输出，允许 C. Neoformans快速微调应力反应，并且此过程中的缺陷与减少有关 病毒。 HOG1 p38 MAP激酶（MAPK）模块是一个介导此过程的信号模块，但是 Hog1介导重编程的机制和效果在Neoformans中未知。我们 提出识别HOG1激活的分子基础的建议，并产生高吞吐量的表征 它的互动组。我们还发现，另一个翻译组调节途径GCN2途径是差异的 在HOG1Δ中受到调节，并旨在评估这两种翻译重新编程中这种变化的含义 以及病毒。这是我们假设HOG1通过其激酶介导的宽阔相互作用的变化 活动，以及与GCN2途径的串扰，在应力耐受性和 Hog1Δ的毒力。我们将测试两个具体目标：（1）确定HOG1调节的机制 Neoformans的应力反应，（2）评估EIF2α磷酸化是否增加了 损失HOG1。该项目很重要，因为它将阐明Hog1重新编程的机理，将会 识别可以针对抑制此过程的效应器，还将检查 两个主要的信号通路。该研究培训计划的总体目标是为候选人配备 回答这些问题的必要技能，并为他们提供相关培训以建立自己 作为研究真菌发病机理和压力适应的成功学术研究者。这些目标将是 在培训的帮助下，在赞助商和论文咨询委员会的指导下完成 大学在布法罗提供的机会和设备，并通过专业发展 会议和研讨会的机会。