The role of histone chaperones and specific transcription factors during multicellular development of filamentous ascomycetes

组蛋白伴侣和特异性转录因子在丝状子囊菌多细胞发育过程中的作用

• 批准号: 240333372
• 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
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/240333372?language=en
• 关键词: role; histone; chaperones; specific; transcription

The aim of this project is the molecular analysis of fruiting body development in filamentous fungi. Fruiting body development is an ontogenetic program that is an essential part of the life cycle of many filamentous fungi; however, its molecular basis is far from understood. In the proposed project we will analyze the role of factors that regulate gene expression at the level of transcription in the model organism Sordaria macrospora. The analyses will be performed with omics-techniques to obtain a genome-wide overview of nuclear processes that occur during fruiting body development. In previous analyses, several transcription factors and histone chaperones involved in fruiting body development were identified. Furthermore, we have established a combination of laser microdissection and RNA-seq to study the organ-specific gene expression in young fruiting bodies of the wild type and the developmental mutant pro1 that carries a mutation in a transcription factor gene. These analyses will be extended to include other transcription factor and histone chaperone mutants to unravel the regulatory network that coordinates chromatin structure and transcription during fruiting body development. In addition, we will establish ChIP-seq to identify genomic regions that are bound by transcription factors. A comparison of RNA-seq and ChIP-seq data can be used to distinguish between direct and indirect targets of transcription factors. We will also include histone chaperones in the ChIP-seq analyses; this requires crosslinking of histone chaperones, histones, and DNA prior to the chromatin immunoprecipitation, because the histone chaperones are not directly binding to DNA. These techniques will be used to analyze the influence of histone chaperones and transcription factors in parallel, which allows the identification of interdependencies. A second aspect of the project is the identification of proteins that interact with the histone chaperones and transcription factors under investigation. We have already identified interaction partners of the histone chaperone ASF1 using a highly sensitive mass spectrometry (MudPIT), and these analyses will be performed for the other developmental factors. The combination of several high-throughput techniques for the analysis of gene expression will yield an integrated picture with a high temporal and spatial resolution of the molecular processes that are orchestrate fruiting body development.

本项目的目的是对丝状真菌子实体发育的分子分析。子实体发育是一个个体发育过程，是许多丝状真菌生活史中必不可少的一部分；然而，其分子基础还远未被了解。在拟议的项目中，我们将分析在模式生物大孢子菌中在转录水平上调节基因表达的因素的作用。这些分析将使用组学技术进行，以获得子实体发育期间发生的核过程的全基因组概述。在以前的分析中，已经鉴定了几个与子实体发育有关的转录因子和组蛋白伴侣蛋白。此外，我们建立了激光显微切割和RNA-SEQ相结合的方法来研究野生型幼子实体和携带转录因子基因突变的发育突变体Pro1中器官特异性基因的表达。这些分析将扩展到包括其他转录因子和组蛋白伴侣突变体，以揭示在子实体发育过程中协调染色质结构和转录的调控网络。此外，我们将建立CHIP-SEQ来识别与转录因子结合的基因组区域。RNA-SEQ和CHIP-SEQ数据的比较可用于区分转录因子的直接靶标和间接靶标。我们还将在芯片序列分析中包括组蛋白伴侣；这需要在染色质免疫沉淀之前交联组蛋白伴侣蛋白、组蛋白和DNA，因为组蛋白伴侣蛋白不直接与DNA结合。这些技术将被用于并行分析组蛋白伴侣和转录因子的影响，从而能够识别相互依赖关系。该项目的第二个方面是鉴定与正在研究的组蛋白伴侣和转录因子相互作用的蛋白质。我们已经使用高灵敏质谱仪(MudPIT)确定了组蛋白伴侣ASF1的相互作用伙伴，并将对其他发育因素进行这些分析。几种用于基因表达分析的高通量技术的组合将产生一幅具有高时间和空间分辨率的综合图像，该图像具有协调子实体发育的分子过程。

项目成果

Innovation and constraint leading to complex multicellularity in the Ascomycota.

• DOI: 10.1038/ncomms14444
• 发表时间: 2017-02-08
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Nguyen TA;Cissé OH;Yun Wong J;Zheng P;Hewitt D;Nowrousian M;Stajich JE;Jedd G
• 通讯作者: Jedd G

New insights from an old mutant: SPADIX4 governs fruiting body development but not hyphal fusion in Sordaria macrospora

• DOI: 10.1007/s00438-016-1258-0
• 发表时间: 2016-10
• 期刊: Molecular Genetics and Genomics
• 影响因子: 3.1
• 作者: I. Teichert;Miriam Lutomski;R. Märker;M. Nowrousian;U. Kück

The transcription factor PRO44 and the histone chaperone ASF1 regulate distinct aspects of multicellular development in the filamentous fungus Sordaria macrospora

• DOI: 10.1186/s12863-018-0702-z
• 发表时间: 2018-12
• 期刊: BMC Genetics
• 影响因子: 2.9
• 作者: David Immanuel Schumacher;Ramona Lütkenhaus;F. Altegoer;I. Teichert;U. Kück;M. Nowrousian