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 新型隐球菌是一种环境真菌，也是患有受损人群的机会性病原体 估计每年导致 181,000 人死亡的免疫系统是新型隐球菌发病机制的关键一步。 是对宿主的适应，这是由几种信号转导途径介导的。 重编程，即蛋白质合成的变化，是这些信号转导途径的关键输出，使得 C. neoformans 能够快速微调应激反应，而该过程中的缺陷与减少 Hog1 p38 MAP 激酶 (MAPK) 模块是介导这一过程的一个信号模块。 Hog1 介导重编程的机制和效应器在新型隐球菌中尚不清楚。 提议确定 Hog1 激活的分子基础，并生成高通量表征 我们还发现另一个翻译组调控途径，Gcn2 途径，是有差异的。 hog1Δ 中调节，旨在评估这种变化对翻译组重编程的影响 因此，我们研究 Hog1 通过其激酶介导广泛的相互作用组的变化。 活性，并且与 Gcn2 通路的串扰掩盖了应激耐受性和 hog1Δ 的毒力我们将测试两个具体目标：（1）确定 Hog1 的调节机制。 新型隐球菌的应激反应，以及 (2) 评估 eIF2α 磷酸化的增加是否可以补偿 对于 Hog1 的丢失，该项目意义重大，因为它将阐明 Hog1 的重编程机制，将 确定可以有针对性地抑制这一过程的效应器，并且还将检查之间的收敛点 该研究培训计划的总体目标是为候选人提供两个主要的信号传导途径。 回答这些问题所需的技能，并为他们提供相关培训以建立自己的能力 作为一名成功的学术研究者，研究真菌发病机制和应激适应。 在赞助商和论文咨询委员会的指导下，在培训的帮助下完成 布法罗大学提供的机会和设备，以及通过专业发展 参加会议和研讨会的机会。