Light-regulated genes affecting the pathogen potential of Cryptococcus neoformans

影响新型隐球菌致病潜力的光调控基因

• 批准号: 7249522
• 负责人: ALEXANDER IDNURM
• 金额: $ 16.14万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7249522
• 关键词: Affect; Area; Cell Extracts; Cell Line; Communicable Diseases; Cryptococcus neoformans; Darkness; Development Plans; Disease; Environment; Exposure to; Gene Expression Regulation; Gene Proteins; Genes; Genetic Transcription; Genome; Goals; Heme; Human; Hypersensitivity; Infection; K22 Award; Laboratories; Light; Link; Mediating; Microarray Analysis; Modeling; Nature; Northern Blotting; Partner in relationship; Phenotype; Photobiology; Photoreceptors; Phototropism; Physiologic pulse; Pigments; Porphyrins; Positioning Attribute; Postdoctoral Fellow; Production; Property; Proteins; Pulse taking; Recombinants; Reproduction spores; Research; Research Personnel; Research Proposals; Resources; Role; Signal Transduction; Testing; UV sensitive; Virulence; Virulent; asexual; career; circadian pacemaker; design; experience; ferrochelatase; fungus; gel mobility shift assay; heme biosynthesis; human disease; macrophage; member; mutant; pathogen; programs; promoter; protoporphyrin IX; transcription factor; ultraviolet irradiation; white collar 1

DESCRIPTION (provided by applicant): The applicant is currently an experienced post-doctoral fellow transitioning into a role as an independent investigator. Immediate career objectives are to complete current research and obtain bridging resources to facilitate the establishment of an academic research position. The K22 award and career development plans would link within the long-term career objectives of developing an internationally-recognized laboratory investigating fungi that cause human disease or allergy. Light influences many properties in species of fungi, such as regulating sexual or asexual sporulation, pigment production, phototropism, and setting a circadian clock. The BWC1 and BWC2 genes, encoding a putative photoreceptor and interacting transcription factor, control inhibition of mating by light, tolerance to UV irradiation, and virulence in the human pathogenic fungus Cryptococcus neoformans. However, it is unclear what genes are regulated by light via Bwc1-Bwc2. Their identification, through this research, will provide directions towards controlling fungal infectious disease. Aim 1 is to identify genes regulated by light using a whole genome microarray analysis of transcription, comparing a wild type strain grown in constant darkness or treated with a pulse of light. Aim 2 is to demonstrate that the promoters of light-regulated genes physically interact with Bwc1-Bwc2 by gel mobility shift assays. Preliminary studies have revealed 30 genes that are light-regulated. One such gene encodes ferrochelatase, which catalyzes the final step of heme biosynthesis from protoporphyrin IX. Heme is an essential co-factor in many proteins, while porphyrins are highly photoreactive. Aim 3 will test the hypothesis that ferrochelatase activity is a major factor that mediates C. neoformans sensitivity to light and virulence. Strains with altered expression levels of ferrochelatase will be constructed and their phenotypes tested for mating, UV sensitivity and virulence in a macrophage cell line model. The relevance of the research is that it will enable the discovery of new genes that enable fungi to be virulent towards humans and to spread in nature. This information is expected to provide strategies to minimize exposure to fungal inocula to reduce initial infection or exposure to spores that cause allergies, and enable the design of strategies to use light as a therapy against fungal diseases.

描述（由申请人提供）：申请人目前是一名经验丰富的博士后研究员，正在过渡到独立研究员的角色。近期的职业目标是完成当前的研究，并获得桥接资源，以促进学术研究职位的建立。K22奖和职业发展计划将与长期职业目标相联系，即建立一个国际公认的实验室，调查导致人类疾病或过敏的真菌。 光影响真菌物种的许多特性，例如调节有性或无性孢子形成、色素产生、向光性和设定生物钟。BWC 1和BWC 2基因，编码一个假定的光感受器和相互作用的转录因子，控制交配的光抑制，耐受紫外线照射，和毒力在人类致病真菌新型隐球菌。然而，目前还不清楚哪些基因通过Bwc 1-Bwc 2受到光的调控。通过这项研究，它们的鉴定将为控制真菌传染病提供方向。 目的1是通过全基因组微阵列转录分析，比较在恒定黑暗中生长或用光脉冲处理的野生型菌株，鉴定受光调控的基因。目的2是通过凝胶迁移率变动分析证明光调控基因的启动子与Bwc 1-Bwc 2的物理相互作用。初步研究已经揭示了30个受光调控的基因。一个这样的基因编码亚铁螯合酶，其催化血红素从原卟啉IX生物合成的最后一步。血红素是许多蛋白质中必不可少的辅因子，而卟啉是高度光反应性的。目的3将验证铁螯合酶活性是介导C.对光线和毒力的敏感性。将构建铁螯合酶表达水平改变的菌株，并在巨噬细胞系模型中检测其交配、UV敏感性和毒力的表型。 这项研究的相关性在于，它将能够发现新的基因，使真菌对人类具有毒性并在自然界中传播。这些信息预计将提供策略，以尽量减少暴露于真菌接种物，以减少初始感染或暴露于孢子，导致过敏，并使策略的设计，使用光作为治疗真菌疾病。