Functional Analysis and Systems Biology of Filamentous Fungi

描述(由申请人提供)：根据ARRA指南NOT-OD-09-058(NIH宣布可为竞争性修订申请提供恢复法资金)提交的补充资金提案扩大了父计划项目拨款的范围。整个努力的中心是粗糙脉孢菌，这是一种超过25万种非酵母真菌的主要丝状真菌模型。一个主要的目标是了解粗毛拟青霉如何从菌丝生长过渡到完全的无性孢子发育。我们把重点放在无性发育的两个关键触发因素上，光和干燥。此外，我们将利用我们之前的成功，将系统淘汰扩展到另一个突出的模型系统，即尼杜拉曲霉。脉孢子菌和曲霉菌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植物病原体以及产生化学品、酶和药物的工业菌株。在之前的5年支持中，我们彻底改变了丝状菌的基因敲除技术 真菌，超过我们最初目标的50%。项目1完成脉孢子菌基因敲除，并将系统性干扰扩展到曲霉菌。为了支持了解管理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补编创建了一个新的曲霉目标，重点是蛋白质组分析，以描述细胞内复合体的组成和位置，同时完成自提交父母拨款以来添加到曲霉基因组中的1242个基因的敲除盒式磁带。在母基金项目#2和#3中，将通过组蛋白修饰、转录因子结合位点和表观遗传标记的ChlP-seq作图，从染色质结构水平通过基因调控网络描述和重建粗毛拟青霉对光和空气的发育反应，将这些与转录组测量相关联，以产生对基因组和表观基因组动态的深入描述。项目3的补编通过纳入两名新的调查员和确定了更多监管者的替代办法，大大扩大了这方面的范围。在ARRA方面，这些补充材料扩大了最初目标的范围，增加了将大大加快工作步伐的项目，这些项目可以在两年内完成。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员来留住员工，聘用一名 失业的技术人员，三名失业的博士后，两名研究生，恢复另一名博士后的充分就业，并大量购买美国制造的用品和设备。 公共卫生进展：丝状真菌，通常被称为霉菌，是常见的动植物病原体，但它们也被广泛用作工业菌株，提供抗生素、化学品、酶和药品。没有他们我们早就死了，但他们会杀了我们。我们试图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好事和控制真菌产生的坏事。这一补充将支持一名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。

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.