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甲基转移酶AflSet1参与真菌形态发生、黄曲霉毒素B1生物合成及毒力。

The Methyltransferase AflSet1 Is Involved in Fungal Morphogenesis, AFB1 Biosynthesis, and Virulence of .

作者信息

Liu Yaju, Zhang Mengjuan, Xie Rui, Zhang Feng, Wang Sen, Pan Xiaohua, Wang Shihua, Zhuang Zhenhong

机构信息

Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China.

出版信息

Front Microbiol. 2020 Feb 18;11:234. doi: 10.3389/fmicb.2020.00234. eCollection 2020.

Abstract

The filament fungal pathogen, , spreads worldwide and contaminates several important crops. Histone posttranslational modifications are deeply involved in fungal development and virulence, but the biological function of the histone methyltransferase AflSet1 in is still unknown. In the study, deletion strain was constructed through homologous recombination, and it was found that AflSet1 up-regulates hyphae growth, and promotes conidiation by sporulation regulation genes: and . It was also found that AflSet1 involves in sclerotia formation and AFB1 biosynthesis via sclerotia related transcriptional factors and orthodox AFB1 synthesis pathway, respectively. Crop models revealed that AflSet1 plays critical roles in colonization and AFB1 production on crop kernels. Lipase activity analysis suggested that AflSet1 affects fungal virulence to crops via digestive enzymes. Stresses tests revealed that AflSet1 is deeply involved in fungal resistance against osmotic, oxidative and cell membrane stress. The preparation of N_SET, SET domain deletion mutants and H988K mutant revealed that both domains play critical roles in fungal development and AFB1 production, and that H988 is very important in executing biological functions on morphogenesis and AFB1 synthesis. Subcellular location analysis revealed that AflSet1 is stably accumulated in nuclei in both spore germination and hyphae growth stages, even under the stress of SDS. Through immunoblot analysis, it was found that AflSet1 methylates H3K4me2 and me3 as well as H3K9me2. This study provides a solid evidence to discover the biological functions of histone methyltransferase in pathogenic fungi.

摘要

丝状真菌病原体……在全球范围内传播并污染多种重要作物。组蛋白翻译后修饰深度参与真菌的发育和毒力,但组蛋白甲基转移酶AflSet1在……中的生物学功能仍不清楚。在该研究中,通过同源重组构建了AflSet1缺失菌株,发现AflSet1上调菌丝生长,并通过孢子形成调控基因……促进分生孢子形成。还发现AflSet1分别通过与菌核相关的转录因子和正统的黄曲霉毒素B1合成途径参与菌核形成和黄曲霉毒素B1生物合成。作物模型表明,AflSet1在作物籽粒的定殖和黄曲霉毒素B1产生中起关键作用。脂肪酶活性分析表明,AflSet1通过消化酶影响真菌对作物的毒力。胁迫试验表明,AflSet1深度参与真菌对渗透、氧化和细胞膜胁迫的抗性。N_SET、SET结构域缺失突变体和H988K突变体的制备表明,这两个结构域在真菌发育和黄曲霉毒素B1产生中均起关键作用,且H988在执行形态发生和黄曲霉毒素B1合成的生物学功能中非常重要。亚细胞定位分析表明,即使在SDS胁迫下,AflSet1在孢子萌发和菌丝生长阶段均稳定积累于细胞核中。通过免疫印迹分析,发现AflSet1使H3K4me2和me3以及H3K9me2发生甲基化。本研究为发现致病真菌中组蛋白甲基转移酶的生物学功能提供了确凿证据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9acb/7040179/c1ba8fa9d66e/fmicb-11-00234-g001.jpg

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