Functional genomics of morphogenesis in filamentous ascomycetes

丝状子囊菌形态发生的功能基因组学

• 批准号: 324272192
• 负责人: Privatdozentin Dr. Minou Nowrousian
• 金额: --
• 依托单位: Lehrstuhl für Molekulare und Zelluläre Botanik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/324272192?language=en
• 关键词: Functional; genomics; morphogenesis; filamentous; ascomycetes

This project aims at identifying molecular mechanisms that regulate fruiting body development in filamentous ascomycetes. During the first phase of the project, we performed comparative transcriptomics analyses of the three ascomycetes Ascodesmis nigricans, Pyronema confluens, and Sordaria macrospora to identify evolutionary conserved patterns of gene expression. The results indicate that gene expression patterns in fruiting bodies are to some degree conserved and are quite distinct from gene expression patterns in vegetative mycelium. Among 83 genes that are upregulated in developing fruiting bodies of all three species, genes encoding proteins with predicted roles in vesicle transport, the endomembrane system, or transport across membranes, and genes encoding proteins with predicted roles in chromatin organization or the regulation of gene expression are strongly represented. Four genes with conserved expression during fruiting body development were functionally characterized by generating deletion mutants in S. macrospora. Thereby, three novel developmental genes were identified. In the proposed second phase of the project, we will focus on genes encoding chromatin modifiers and transcription factors, and their role in developmental regulation of chromatin structure, histone modifications and gene expression. Preliminary data indicate that histone modifications might be different from the wild type in the S. macrospora histone chaperone mutant asf1, which has a block during early fruiting body development. We will analyze histone modifications in the S. macrospora wild type and developmental mutants at a genome-wide level using ChIP-seq. Modifications that change during development will also be analyzed in different developmental stages in A. nigricans and P. confluens. Furthermore, we will analyze the chromatin structure at different developmental stages using Hi-C, and analyze the results together with transcriptome data and results on histone modifications to establish an integrated picture of chromatin and gene expression changes during fungal development. In addition, we will analyze protein-protein interactions between chromatin modifier proteins, identify protein domains required for these interactions and for the developmental functions of the proteins, and characterize additional putative developmental genes based on the results of available and novel gene expression data.

本研究旨在探讨丝状子囊菌子实体发育的分子调控机制。在该项目的第一阶段，我们进行了比较转录组学分析的三个子囊菌Ascodesmis nigricans，Pyronema confluens，和Sordaria macrospora，以确定基因表达的进化保守模式。结果表明，子实体中的基因表达模式具有一定的保守性，与营养菌丝体中的基因表达模式截然不同。在所有三个物种的子实体发育中上调的83个基因中，编码在囊泡运输、内膜系统或跨膜运输中具有预测作用的蛋白质的基因，以及编码在染色质组织或基因表达调控中具有预测作用的蛋白质的基因被强烈地代表。通过构建缺失突变体，对4个在子实体发育过程中具有保守表达的基因进行功能鉴定。大孢子虫。由此，鉴定出三个新的发育基因。在该项目的第二阶段，我们将专注于编码染色质修饰剂和转录因子的基因，以及它们在染色质结构，组蛋白修饰和基因表达的发育调控中的作用。初步数据表明，组蛋白修饰可能不同于野生型的S。macrospora组蛋白分子伴侣突变体p5 asf 1在早期子实体发育过程中具有阻滞作用。我们将分析S.大孢子菌野生型和发育突变体在全基因组水平使用ChIP-seq.在发育过程中发生变化的修饰也将在A. nigricans和P. confluens。此外，我们将使用Hi-C分析不同发育阶段的染色质结构，并将结果与转录组数据和组蛋白修饰结果一起分析，以建立真菌发育过程中染色质和基因表达变化的综合图像。此外，我们将分析染色质修饰蛋白之间的蛋白质-蛋白质相互作用，确定这些相互作用和蛋白质的发育功能所需的蛋白质结构域，并根据现有的和新的基因表达数据的结果表征其他推定的发育基因。