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宣布恢复法案资金用于竞争性修订申请的可用性）提交的补充资金提案扩展了母计划项目赠款的范围。整体工作集中在粗糙脉孢菌上，这是一种超过25万种非酵母真菌的首要丝状真菌模型。一个主要的目标是了解如何N。crassa从菌丝生长过渡到完全的无性孢子发育。我们专注于无性发育的两个关键触发因素，光照和干燥。此外，我们将利用我们以前的成功，扩大系统敲除到一个额外的突出的模型系统，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。在前5年的前期支持中，我们彻底改变了丝状菌基因敲除技术， 真菌，超过我们最初的目标> 50%。项目#1完成脉孢菌基因敲除并将系统性破坏扩展到曲霉菌。为了支持理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，通过高通量技术产生敲除和敲入。该项目#1补充创建了一个新的曲霉目标，专注于蛋白质组学分析，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基因组中的1242个基因的敲除盒。在母基金项目#2和#3中，将描述和重建N. crassa对光和空气的发育反应，从染色质结构水平通过基因调控网络，通过ChIP-seq映射组蛋白修饰，转录因子结合位点和表观遗传标记，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的补充大大扩大了这一范围，纳入了两名新的调查人员和一种确定了其他监管机构的替代方法。在ARRA方面，补充文件通过增加项目来扩大最初目标的范围，这些项目将大大加快工作的步伐，并可以在2年内完成。它们允许创造就业机会 通过增加一名尚未得到NIH支持的初级教师新研究员， 失业的技术人员，三个失业的博士后，两个研究生，恢复充分就业的另一个博士后，并作出大量购买美国制造的用品和设备。 公共卫生部门：丝状真菌，通常被称为霉菌，是常见的动物和植物病原体，但它们也被广泛用作工业菌株，以提供抗生素，化学品，酶和药物。没有他们我们早就死了但他们能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东西并控制坏东西。该补充将支持NIH新的研究员，并雇用或恢复充分就业的技术人员，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.