Abstract:
Lysine acetylation is an evolutionarily conserved and widespread post-translational modification implicated in the regulation of multiple metabolic processes, but its function remains largely unknown in plant pathogenic fungi. A comprehensive analysis combined with proteomic, molecular and cellular approaches was presented to explore the roles of cytoplasmic acetylation in Fusarium oxsysporum f.sp. lycopersici (Fol). The divergent cytoplasmic deacetylase FolSir2 was biochemically characterized, which is contributing to fungal virulence. Based on this, a total of 1752 acetylated sites in 897 proteins were identified in Fol via LC-MS/MS analysis. Further analyses of the quantitative acetylome revealed that 115 proteins representing two major pathways, translational and ribosome biogenesis, were hyperacetylated in the ∆Folsir2 strain. We experimentally examined the regulatory roles of FolSir2 on K271 deacetylation of FolGsk3, a serine/tyrosine kinase implicated in a variety of cellular functions, which was found to be crucial for the activation of FolGsk3 and thus modulated Fol pathogenicity. Cytoplasmic deacetylation by FolSir2 homologues has a similar function in Botrytis cinerea and likely other fungal pathogens. These findings reveal a conserved mechanism of silent information regulator 2-mediated cytoplasmic deacetylation that is involved in plant-fungal pathogenicity, providing a candidate target for designing broad-spectrum fungicides to control plant diseases.
摘要:
赖氨酸乙酰化是一种进化上保守且广泛的翻译后修饰,涉及多种代谢过程的调节。但它的功能在植物病原真菌中仍然未知。结合蛋白质组学的综合分析,提出了分子和细胞方法来探索细胞质乙酰化在镰刀菌中的作用。lycopersici(Fol)。不同的细胞质去乙酰化酶FolSir2进行了生化表征,这有助于真菌的毒力。基于此,通过LC-MS/MS分析在Fol中鉴定出897种蛋白质中的总共1752个乙酰化位点。进一步的定量分析显示,115个蛋白质代表两个主要途径,翻译和核糖体生物发生,在ΔFolsir2菌株中高度乙酰化。我们通过实验检查了FolSir2对FolGsk3的K271脱乙酰化的调节作用,FolGsk3是一种涉及多种细胞功能的丝氨酸/酪氨酸激酶,发现这对于FolGsk3的激活至关重要,因此可以调节Fol的致病性。通过FolSir2同源物进行的细胞质去乙酰化在灰葡萄孢和可能的其他真菌病原体中具有相似的功能。这些发现揭示了沉默信息调节因子2介导的细胞质去乙酰化的保守机制,该机制与植物真菌的致病性有关。为设计广谱杀菌剂控制植物病害提供了候选靶标。