禾谷镰孢<span>Rpd3</span><span>复合体亚基</span><span>Sin3</span><span>的错义突变回复</span><i><span>fng1</span></i>突变体缺陷的机制

Journal of Integrative Agriculture

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禾谷镰孢Rpd3复合体亚基Sin3的错义突变回复fng1突变体缺陷的机制

修回日期:2024-02-01

A missense mutation in the Sin3 subunit of Rpd3 histone deacetylase complex bypasses the requirement for FNG1 in wheat scab fungus

Xu Huai-jian¹, Jiang Ruo-xuan¹, Fu Xian-hui¹, Wang Qin-hu¹, Shi Yu-tong¹, Zhao Xiao-fei¹, Jiang Cong^1#, Jiang Hang^2#

¹ State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling 712100, China
²Shandong Key Laboratory of Plant Virology, Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan 250100, China

Revised:2024-02-01
About author:Xu Huai-jian, E-mail: huaijianxu@yeah.net; #Correspondence Jiang Cong, Tel: +86-153-3903-4156, E-mail: cjiang@nwafu.edu.cn; Jiang Han, Tel: +86-173-2935-3665, E-mail: jhfor724@163.com

摘要/Abstract

摘要： 由禾谷镰孢（Fusarium graminearum）引发的小麦赤霉病是一种重要的农作物真菌病害。禾谷镰孢不仅能够侵染小麦，造成产量降低，还会产生多种毒素，威胁人畜健康，对食品安全构成巨大威胁。禾谷镰孢整个生活史，包括生长、致病、产毒及有性生殖等生命活动都受到了组蛋白乙酰化等修饰的调控。组蛋白乙酰化状态由组蛋白乙酰化复合体（HAT）和去乙酰化复合体（HDAC）共同调节。其中Rpd3复合体作为最重要的组蛋白去乙酰化复合体之一而受到广泛关注。Sin3是Rpd3复合体中最大的亚基，对其关键功能位点及作用机制尚缺乏认识。在本研究中，我们鉴定到位于第810位的谷氨酸残基（E810）是禾谷镰孢FgSin3上的关键位点。研究发现，FgSin3的E810位点突变会引起禾谷镰孢菌丝生长、有性生殖、致病力及DON毒素合成的缺陷。此外，E810K的错义突变能够提高因FNG1（人类生长抑制基因ING1的同源基因）缺失而降低的H4乙酰化水平，从而部分回复fng1突变体的表型缺陷。经序列比对和进化分析，发现FgSin3的同源基因在真核生物中普遍存在，并且其E810功能位点在真菌、动物和植物中表现出高度保守性。基于Alphafold2预测的结构，E810位于FgRpd3-FgSin3的互作界面上，并与FgRpd3的R317位点形成氢键。E810的模拟突变会破坏该氢键的形成，进而影响FgRpd3和FgSin3间的蛋白互作。综上，FgSin3的E810位点可能通过影响Rpd3复合体亚基的组装，与Fng1共同维系真菌组蛋白H4乙酰化平衡，并介导复杂的生物学过程。鉴于FgSin3功能及序列的保守性，以及E810位点周围残基在不同物种中的偏好性差异，未来有望作为药物研发的候选靶标。本研究鉴定到Sin3中的关键功能位点，并从生物学功能、结构基础和进化模式等方面对其参与Rpd3复合体组装的机制和途径进行了解析。

Abstract: The Rpd3 histone deacetylase complex is a multiple-subunit complex that mediates the regulation of chromatin accessibility and gene expression. Sin3, the largest subunit of Rpd3 complex, is conserved in a broad range of eukaryotes. Despite being a molecular scaffold for complex assembly, the functional sites and mechanism of action of Sin3 remain unexplored. In this study, we functionally characterized a glutamate residue (E810) in FgSin3, the ortholog of yeast Sin3 in Fusarium graminearum (known as wheat scab fungus). Our findings indicate that E810 was important for the functions of FgSin3 in regulating vegetative growth, sexual reproduction, wheat infection, and DON biosynthesis. Furthermore, the E810K missense mutation restored the reduced H4 acetylation caused by the deletion of FNG1, the ortholog of the human inhibitor of growth (ING1) gene in F. graminearum. Correspondingly, the defects of the fng1 mutant were also partially rescued by the E810K mutation in FgSin3. Sequence alignment and evolutionary analysis revealed that E810 residue is well-conserved in fungi, animals, and plants. Based on Alphafold2 structure modeling, E810 localized on the FgRpd3-FgSin3 interface for the formation of a hydrogen bond with FgRpd3. Mutation of E810 disrupts the hydrogen bond and likely affects the FgRpd3-FgSin3 interaction. Taken together, E810 of FgSin3 is functionally associated with Fng1 in the regulation of H4 acetylation and related biological processes, probably by affecting the assembly of the Rpd3 complex.

Key words: Histone acetylation , ING protein , , Phytopathogen , , Rpd3 histone deacetylase complex

. 禾谷镰孢Rpd3复合体亚基Sin3的错义突变回复fng1突变体缺陷的机制[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.01.006.

Xu Huai-jian, Jiang Ruo-xuan, Fu Xian-hui, Wang Qin-hu, Shi Yu-tong, Zhao Xiao-fei, Jiang Cong, Jiang Hang. A missense mutation in the Sin3 subunit of Rpd3 histone deacetylase complex bypasses the requirement for FNG1 in wheat scab fungus[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.01.006.

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