Functional Analysis and Systems Biology of Filamentous Fungi

丝状真菌的功能分析和系统生物学

• 批准号: 7814793
• 负责人: Jay C. Dunlap
• 金额: $ 134.54万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7814793
• 关键词: Administrative Supplement; Air; Ally; Animals; Antibiotics; Aspergillus; Aspergillus nidulans; Basic Science; Binding Sites; Biological; Biological Models; Biology; Cells; Chemicals; Chromatin; Chromatin Structure; Classification; Communities; Complex; Desiccation; Development; Electronic Mail; Employment; Enzymes; Epigenetic Process; Equipment and Supplies; Eukaryota; Exposure to; Faculty; Fluorescence Microscopy; Funding; Fungi Model; Gene Expression Profile; Gene Proteins; Genes; Genome; Genomics; Goals; Grant; Growth; Growth and Development function; Guidelines; Knock-in Mouse; Knock-out; Light; Location; Maps; Measurement; Modeling; Molds; National Institute of General Medical Sciences; Neurospora; Neurospora crassa; Occupations; Ohio; Oligonucleotides; Parents; Pharmacologic Substance; Plants; Postdoctoral Fellow; Program Research Project Grants; Proteomics; Public Health; Recovery; Regulator Genes; Regulatory Pathway; Reproduction spores; Research; Research Infrastructure; Research Personnel; Seasons; Series; Students; Systems Biology; Techniques; Technology; Unemployment; United States National Institutes of Health; Universities; Work; Writing; Yeasts; asexual; base; fundamental research; fungal genetics; fungus; genome-wide; graduate student; histone modification; killings; knockout gene; parent grant; pathogen; programs; protein protein interaction; response; success; tool; transcription factor

DESCRIPTION (provided by applicant): This proposal for supplementary funding, submitted under ARRA guidelines NOT-OD-09-058 (NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications) extends the scope of the parent Program Project grant. The overall effort centers on Neurospora crassa, a premier filamentous fungus model for over 250,000 species of non-yeast fungi. A primary goal is to understand how N. crassa transitions from mycelial growth to complete asexual spore development. We focus on two key triggers of asexual development, light and desiccation. In addition, we will leverage our prior successes to expand systematic knockouts to an additional prominent model system, Aspergillus nidulans. Neurospora and Aspergillus are salient models for basic research in eukaryotes; fungi allied to these species include most animal and plant pathogens as well as industrial strains yielding chemicals, enzymes, and pharmaceuticals. Over the first 5 years of prior support we revolutionized the technology for gene knockouts in filamentous fungi, exceeding our initial target by >50%. Project #1 completes Neurospora gene knockouts and extends systematic disruptions to Aspergillus. In support of the overarching goal of understanding regulatory pathways governing filamentous fungal development, in Projects #2 and #3, knockout and knock-ins are created via high throughput techniques. The Project #1 Supplement creates a new Aspergillus aim focused on proteomic analyses to describe the composition and location of intracellular complexes, while completing knockout cassettes for 1242 genes added to the Aspergillus genome since the parent grant was submitted. In the parent grant Projects #2 and #3 will describe and reconstruct the regulatory cascade that underlies N. crassa's developmental response to light and air, from the level of chromatin structure through the gene regulatory network, via ChlP-seq mapping of histone modifications, transcription factor binding sites, and epigenetic marks, correlating these with transcriptome measurements to generate a deep description of genome and epigenome dynamics. The Supplement to Project #3 expands the scope of this considerably by incorporating two new investigators and an alternative approach that has identified additional regulators. In terms of the ARRA, the supplements expand the scope of the original aims by adding projects that will vastly accelerate the pace ofthe work and that can be completed within 2 years. They allow for job creation and retention by adding a junior faculty New Investigator not yet supported by the NIH, employing an unemployed technician, three unemployed postdocs, two graduate students, restoring to full employment another postdoc, and making substantial purchases of US-made supplies and equipment. PUBLIC HEALTH REVELANCE: Filamentous fungi, typically known as molds, are common animal and plant pathogens, but they are also widely used as industrial strains to provide antibiotics, chemicals, enzymes, and pharmaceuticals. We'd be dead without them but they can kill us. We seek to understand how genes and proteins work together to regulate fungal growth and development, so as to enhance the good things and control the bad things produced by fungi. This supplement would support an NIH New Investigator, and employ or restore to full employment a technician, four postdocs, and two students.

说明（由申请人提供）：根据 ARRA 指南 NOT-OD-09-058（NIH 宣布用于竞争性修订申请的恢复法案资金的可用性）提交的补充资金提案扩展了父计划项目拨款的范围。总体工作集中在粗糙脉孢菌（Neurospora crassa）上，这是一种适用于超过 250,000 种非酵母真菌的首要丝状真菌模型。主要目标是了解粗糙链球菌如何从菌丝生长过渡到完全无性孢子发育。我们关注无性发育的两个关键触发因素：光照和干燥。此外，我们将利用之前的成功经验，将系统敲除扩展到另一个著名的模型系统——构巢曲霉。脉孢菌属和曲霉属是真核生物基础研究的重要模型；与这些物种相关的真菌包括大多数动物和植物病原体以及产生化学品、酶和药物的工业菌株。在先前支持的前 5 年里，我们彻底改变了丝状基因敲除技术 真菌，超过我们最初的目标 50% 以上。项目#1 完成了脉孢菌基因敲除，并将系统性破坏扩展到曲霉属。为了支持了解控制丝状真菌发育的调控途径的总体目标，在项目#2和#3中，通过高通量技术创建了敲除和敲入。项目 #1 补充文件创建了一个新的曲霉属目标，重点是蛋白质组分析，以描述细胞内复合物的组成和位置，同时完成自提交母基金以来添加到曲霉属基因组中的 1242 个基因的敲除盒。在母基金项目#2和#3中，将通过组蛋白修饰的 ChlP-seq 作图，从染色质结构水平到基因调控网络，描述和重建粗糙猪笼草对光和空气发育反应的调控级联，转录因子结合位点和表观遗传标记，将它们与转录组测量相关联，以生成基因组和表观基因组动态的深入描述。项目 #3 的补充通过纳入两名新的调查员和已确定其他监管机构的替代方法，大大扩展了其范围。就 ARRA 而言，补充文件通过增加项目扩大了最初目标的范围，这些项目将大大加快工作节奏，并且可以在 2 年内完成。它们可以创造就业机会 并通过增加一名尚未得到 NIH 支持的初级教员新研究者、聘用一名 失业的技术人员、三名失业的博士后、两名研究生、另一名博士后恢复充分就业，并大量购买美国制造的用品和设备。 公共卫生启示：丝状真菌（通常称为霉菌）是常见的动植物病原体，但它们也广泛用作工业菌株来提供抗生素、化学品、酶和药物。没有他们我们就会死，但他们可以杀了我们。我们寻求了解基因和蛋白质如何共同作用来调节真菌的生长和发育，从而增强真菌产生的有益物质并控制真菌产生的有害物质。该补助将支持一名 NIH 新研究员，并雇用或恢复充分就业一名技术员、四名博士后和两名学生。