Analysis of programmed cell death in filamentous fungi

丝状真菌程序性细胞死亡分析

• 批准号: 6986582
• 负责人: N Louise GLASS
• 金额: $ 31.64万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6986582
• 关键词: Neurospora; apoptosis; fungal antigens; fungal proteins; heterokaryon; immunity; mitogen activated protein kinase; protein protein interaction

DESCRIPTION (provided by applicant): The primary goal of the proposed studies genetically dissect the mechanism of nonself recognition in filamentous fungi which leads to fungal programmed cell death (PCD) using Neurospora crassa as a model system. Nonself recognition is essential for defense against pathogen invasion and prevention of resource plundering. Our hypothesis is that filamentous fungi, such as Neurospora crassa, have harnessed protein heterocomplex formation as a mechanism to accomplish nonself recognition, which leads to fungal PCD. Nonself recognition during vegetative growth in filamentous fungi is mediated by genetic loci, termed het (for heterokaryon) loci, which regulate the viability of heterokaryons that occur as a result of hyphal fusion between genetically different individuals, het loci show evidence of balancing selection, similar to loci within the Major Histocompatibility Complex (MHC) which are part of the mammalian immune system. Heterokaryon incompatibility is a form of programmed cell death, which can be defined as an orderly process in which a cell actively participates in its own death. Heterokaryon incompatibility is a fungal "immune" system that confers selective advantages to filamentous fungi by preventing hyphal fusion events that spread mycoviruses, debilitated organelles and deleterious plasmids throughout a fungal population. Our initial results (supported by our previous NIH grant) show that nonself recognition occurs by formation of a HET-C protein heterocomplex associated with the plasma membrane, which appears to be fundamentally different in stability/structure than HET-C monomers. Our data suggests that HET-C interacts with a second protein carrying a HET domain; HET domain genes are common, but of unknown function, in filamentous fungal genomes, including human fungal pathogens. This protein complex formation activates signal transduction mechanisms and gene regulation, ultimately resulting in death and destruction of the heterokaryotic cell. The experiments described in this proposal aim to further investigate the role the HET-C/VIB-2 heterocomplex in nonself recognition, and MAP kinase pathways and the VIB-1 transcriptional regulator in triggering fungal PCD. We believe that a dissection of fungal PCD in N. crassa will illuminate conserved and divergent features of PCD among eukaryotic organisms, provide potential drug targets for pathogenic filamentous fungi.

描述（由申请方提供）：拟定研究的主要目标是使用粗糙脉孢菌作为模型系统，从遗传学角度剖析丝状真菌中导致真菌程序性细胞死亡（PCD）的非自我识别机制。非自我识别对于防御病原体入侵和防止资源掠夺是必不可少的。我们的假设是，丝状真菌，如粗糙脉孢菌，利用蛋白质杂合物的形成作为一种机制来完成非自我识别，这导致真菌PCD。丝状真菌营养生长期间的非自我识别由遗传基因座介导，称为het（异核体）基因座，其调节由于遗传上不同的个体之间的菌丝融合而发生的异核体的活力。het基因座显示出平衡选择的证据，类似于作为哺乳动物免疫系统的一部分的主要组织相容性复合物（MHC）内的基因座。异核体不相容性是细胞程序性死亡的一种形式，它可以被定义为细胞主动参与自身死亡的有序过程。异核体不相容性是一种真菌“免疫”系统，其通过阻止菌丝融合事件而赋予丝状真菌选择性优势，菌丝融合事件在整个真菌群体中传播真菌病毒、衰弱的细胞器和有害质粒。我们的初步结果（由我们以前的NIH资助支持）表明，非自我识别是通过形成与质膜相关的HET-C蛋白杂合物而发生的，该杂合物在稳定性/结构上与HET-C单体似乎根本不同。我们的数据表明，HET-C与携带HET结构域的第二种蛋白质相互作用; HET结构域基因在丝状真菌基因组（包括人类真菌病原体）中很常见，但功能未知。这种蛋白质复合物的形成激活信号转导机制和基因调控，最终导致异源细胞的死亡和破坏。本实验旨在进一步研究HET-C/VIB-2异源复合物在非自我识别中的作用，以及MAP激酶途径和VIB-1转录调节因子在触发真菌PCD中的作用。我们认为，解剖真菌PCD在N。crassa将阐明真核生物中PCD的保守性和差异性，为致病丝状真菌提供潜在的药物靶标。