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一个含有 SET 结构域的蛋白质,参与细胞壁完整性信号和过氧化物酶体生物发生,对于炭疽菌分生孢子器的形成和致病性是必需的。

A SET domain-containing protein involved in cell wall integrity signaling and peroxisome biogenesis is essential for appressorium formation and pathogenicity of Colletotrichum gloeosporioides.

机构信息

Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Germplasm Resources Utilization), Ministry of Agriculture, Xingcheng 125100, China.

The Sainsbury Laboratory, University of East Anglia, Norwich Research Park NR4 7UH, UK.

出版信息

Fungal Genet Biol. 2020 Dec;145:103474. doi: 10.1016/j.fgb.2020.103474. Epub 2020 Sep 30.

Abstract

The chromatin modulator Set5 plays important regulatory roles in both cell growth and stress responses of Saccharomyces cerevisiae. However, its function in filamentous fungi remains poorly understood. Here, we report the pathogenicity-related gene CgSET5 discovered in a T-DNA insertional mutant M285 of Colletotrichum gloeosporioides. Bioinformatic analysis revealed that CgSET5 encodes a SET domain-containing protein that is a homolog of the budding yeast S. cerevisiae Set5. CgSET5 is important for hyphae growth and conidiation and is necessary for appressorium formation and pathogenicity. CgSet5 regulates appressorium formation in a mitogen-activated protein kinase-independent manner. Inactivation of CgSET5 resulted in a significant reduction in chitin content within the cell wall, indicating CgSet5 plays a vital role in cell wall integrity. CgSet5 is involved in peroxisome biogenesis. We identified CgSet5 as the histone H4 methyltransferase, which methylates the critical H4 lysine residues 5 and 8 in C. gloeosporioides. We carried out a yeast two-hybrid screen to find CgSet5 interacting partners. We found CgSet5 putatively interacts with an inorganic pyrophosphatase named CgPpa1, which co-localized in the cytoplasm with CgSet5. Finally, CgPpa1 was found to strongly interact with CgSet5 in vivo during appressorium formation by bimolecular fluorescence complementation assays. These data corroborate a complex control function of CgSet5 acting as a core pathogenic regulator, which connects cell wall integrity and peroxisome biogenesis in C. gloeosporioides.

摘要

组蛋白修饰酶 Set5 在酿酒酵母的细胞生长和应激反应中发挥重要的调控作用。然而,其在丝状真菌中的功能尚不清楚。本研究在炭疽菌 T-DNA 插入突变体 M285 中发现了一个与致病性相关的基因 CgSET5。生物信息学分析表明,CgSET5 编码一个 SET 结构域蛋白,是酿酒酵母 Set5 的同源物。CgSET5 对于菌丝生长和产孢至关重要,也是附着胞形成和致病性所必需的。CgSet5 以丝裂原活化蛋白激酶非依赖性的方式调控附着胞的形成。CgSET5 的失活导致细胞壁中几丁质含量显著减少,表明 CgSet5 在细胞壁完整性中发挥着重要作用。CgSet5 参与过氧化物酶体的生物发生。我们鉴定出 CgSet5 是组蛋白 H4 甲基转移酶,它甲基化炭疽菌中的关键 H4 赖氨酸残基 5 和 8。我们进行了酵母双杂交筛选以寻找 CgSet5 的相互作用伙伴。我们发现 CgSet5 可能与一种无机焦磷酸酶 CgPpa1 相互作用,该酶与 CgSet5 在细胞质中共定位。最后,通过双分子荧光互补测定发现,CgPpa1 在附着胞形成过程中与 CgSet5 体内强烈相互作用。这些数据证实了 CgSet5 作为一个核心致病性调控因子的复杂调控功能,它连接了炭疽菌中的细胞壁完整性和过氧化物酶体生物发生。

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