Discerning the roles of novel glycosyltransferases in the pathogenic yeast Cryptococcus neoformans

识别新型糖基转移酶在致病性酵母新型隐球菌中的作用

基本信息

批准号：
10356130
负责人：
Liza Loza
金额：
$ 3.27万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10356130
关键词：
Academia Affinity Chromatography Anabolism Anions Antifungal Agents Biochemical Biological Biological Assay Biology Cell Wall Cells Chromatography Clustered Regularly Interspaced Short Palindromic Repeats Collection Complement Complex Coupled Cryptococcus Cryptococcus neoformans Defect Detection Disease Drug Targeting Enzymes Future Gene Deletion Glycobiology Glycosyltransferase Gene Goals Immunocompromised Host In Vitro Individual National Research Service Award Infection Lung Mediating Meningeal Methods Microbiology Mission Modeling Molecular Mus Nickel Organism Pathogenicity Pathway interactions Performance Persons Pharmaceutical Preparations Phenotype Physiologic pulse Polysaccharides Protein Biochemistry Proteins Publications Reaction Recombinants Research Role Scientist Series Stress Structure Testing Training Virulence Virulence Factors Virulent Work Yeasts capsule career fungus glycosyltransferase human pathogen in vitro Assay innovation interest mortality mutant new therapeutic target novel pathogen pathogenic fungus pre-doctoral response skills sugar nucleotide symposium training opportunity

项目摘要

PROJECT SUMMARY/ABSTRACT Cryptococcus neoformans is an opportunistic fungal pathogen with a high mortality rate. Its major virulence factors are primarily composed of well-characterized glycans; however, the enzymatic steps involved in synthesizing these glycans are largely unknown. The long-term objective of the proposed research is to define unknown steps in glycan synthesis pathways of this organism, mediated by glycosyltransferase enzymes (GTs), that are important for virulence. I propose two complementary approaches to achieve this objective. First, the virulence of GT gene deletion mutants will be compared to that of wild-type C. neoformans to prioritize candidates for study. Mutants lacking GTs of greatest interest will be assessed phenotypically, to suggest biological roles of the GTs, and their glycans analyzed for defects, to suggest specific biochemical roles of the GTs. In parallel, GTs of interest will be recombinantly expressed and their enzymatic activity determined using novel biochemical assays. Together, these approaches will define the roles of GTs that are important in virulence, contributing to our understanding of the basic biology of this pathogen and potentially revealing novel drug targets. Through the successful completion of the proposed research, together with additional formal and informal training, the PI will become technically adept in molecular microbiology, glycobiology, and protein biochemistry. In addition, the PI will cultivate professional skills tailored to a future career in academia by presenting original work at seminars, conferences, and in written form for publication. By facilitating her participation in these predoctoral training opportunities, the Ruth L. Kirschstein Predoctoral Individual National Research Service Award will enable the PI to develop as an independent scientist, in keeping with its mission.

项目概要/摘要新型隐球菌是一种机会性真菌病原菌，死亡率很高。其主要毒力因子主要由特征明确的聚糖组成；然而，涉及的酶促步骤这些聚糖的合成过程在很大程度上是未知的。拟议研究的长期目标是定义该生物体的聚糖合成途径中的未知步骤，由糖基转移酶（GT）介导，这对于毒力很重要。我提出两种互补的方法来实现这一目标。首先， GT 基因缺失突变体的毒力将与野生型新型隐球菌的毒力进行比较，以确定候选者的优先顺序用于学习。缺乏最感兴趣的 GT 的突变体将进行表型评估，以表明其生物学作用 GT 的结构及其聚糖缺陷分析，以表明 GT 的特定生化作用。并联，感兴趣的 GT 将被重组表达，并使用新型生化方法测定其酶活性化验。这些方法共同将定义 GT 在毒力方面的重要作用，有助于我们对这种病原体的基本生物学的了解以及可能揭示的新药物靶点。通过成功完成拟议的研究以及额外的正式和非正式培训后，PI 将精通分子微生物学、糖生物学和蛋白质生物化学。此外，PI 将通过在研讨会上展示原创作品来培养适合未来学术界职业的专业技能，会议，并以书面形式出版。通过促进她参与这些博士前培训机会，Ruth L. Kirschstein 博士前个人国家研究服务奖将使 PI 发展成为一名独立科学家，履行其使命。