蛋白质组分析揭示了稻瘟病菌延伸复合物蛋白PoElp<span>3</span><span>通过调节</span><span>tRNA</span><span>介导的翻译效率影响发育、致病性和自噬的机制</span>

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (4): 1515-1528.DOI: 10.1016/j.jia.2024.01.027

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蛋白质组分析揭示了稻瘟病菌延伸复合物蛋白PoElp3通过调节tRNA介导的翻译效率影响发育、致病性和自噬的机制

收稿日期:2023-04-29 接受日期:2023-12-22 出版日期:2025-04-20 发布日期:2025-03-18

Proteomic analysis revealed the function of PoElp3 in development, pathogenicity, and autophagy through the tRNA-mediated translation efficiency in the rice blast fungus

Yuanhao Liu^1*, Ting Sun^1*, Yuyong Li^1*, Jianqiang Huang^2*, Xianjun Wang², Huimin Bai², Jiayi Hu², Zifan Zhang¹, Shuai Wang¹, Dongmei Zhang², Xiuxiu Li², Zonghua Wang^3#, Huakun Zheng^1#, Guifang Lin^4#

¹Fujian Universities Key Laboratory for Plant-Microbe Interaction, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
²State Key Laboratory of Agricultural and Forestry Biosecurity, College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China
³Fuzhou Institute of Oceanography, Minjiang University, Fuzhou 350108, China
⁴Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China

Received:2023-04-29 Accepted:2023-12-22 Online:2025-04-20 Published:2025-03-18
About author:#Correspondence Guifang Lin, E-mail: glin2150@163.com; Huakun Zheng, E-mail: huakunz2017@fafu.edu.cn; Zonghua Wang, E-mail: wangzh@fafu.edu.cn * These authors contributed equally to this work.
Supported by:
This work was supported by National Natural Science Foundation of China (32172365 and 32272513), the Central Guidance on Local Science and Technology Development Fund of Fujian Province, China (2022L3088), and the Innovative Research Funding of Fujian Agriculture and Forestry University, China (CXZX2020153D).

摘要/Abstract

摘要：

延伸复合物广泛存在于不同物种中，并在许多细胞学过程中发挥重要作用。我们此前的研究结果表明稻瘟病菌延伸复合物蛋白PoElp3参与调控生长发育、致病性和自噬。但其调控机制仍不清楚。为此，本文结合蛋白质组、转录组和定点整合等方法对PoElp3的作用机制进行研究。表型分析结果显示，过表达tK(UUU)和tQ(UUG)两种tRNA可以回补DPoelp3菌株的缺陷，表明PoElp3可能通过调控tRNA介导的蛋白翻译效率发挥作用。在此基础上，我们结合TMT蛋白质组和转录组分析，发现DPoelp3菌株中存在386个丰度较低、但转录水平与野生型菌株Guy11相比无明显差异的蛋白，将其命名为Erp（PoElp3 regulated protein）蛋白。密码子使用偏好性分析结果显示，与70-15全基因组蛋白相比，这386个Erp蛋白的mRNA中具有CAA偏好性的比例较高。除了6个已报道的参与调控生长发育、致病性和自噬的Erp蛋白，我们发现DPoelp3菌株中鞘脂C9-甲基转移酶PoErp9的丰度也降低了。为了检测PoErp9是否只在翻译水平受PoElp3的调控，我们分别在野生型和DPoelp3菌株的ILV2位点敲入PoERP9-GFP融合片段，以规避不同基因组环境对融合片段转录水平的影响。分析结果显示，与野生型相比，突变体中PoErp9-GFP丰度降低，但PoERP9转录本无显著变化，表明PoErp9确实只在翻译水平受PoElp3调控。表型分析结果表明，PoErp9同样在稻瘟病菌菌丝生长、产孢、致病性、以及TOR相关的自噬平衡方面都具有重要的作用。综上所述，我们的研究揭示了PoElp3通过维系tRNA介导的鞘脂代谢途径重要蛋白的翻译效率，进而调控稻瘟病菌生长发育、致病性和自噬的机制。

Abstract:

The Elongator complex is conserved in a wide range of species and plays crucial roles in diverse cellular processes. We have previously shown that the Elongator protein PoElp3 was involved in the asexual development, pathogenicity, and autophagy of the rice blast fungus. In this study, we further revealed that PoElp3 functions via tRNA-mediated protein integrity. Phenotypic analyses revealed that overexpression of two of the tRNAs, tK(UUU) and tQ(UUG) could rescue the defects in ΔPoelp3 strain. TMT-based proteomic and transcriptional analyses demonstrated that 386 proteins were down-regulated in ΔPoelp3 strain compared with wild type strain Guy11, in a transcription-independent manner. Codon usage assays revealed an enrichment of Glutamine CAA-biased mRNA in the 386 proteins compared with the 70-15 genome. In addition to those reported previously, we also found that PoErp9, a sphingolipid C9-methyltransferase, was down-regulated in the ΔPoelp3 strain. Through an ILV2-specific integration of PoERP9-GFP into the wild type and ΔPoelp3 strain, we were able to show that PoErp9 was positively regulated by PoElp3 translationally but not transcriptionally. Functional analyses revealed that PoErp9 was involved in the fungal growth, conidial development, pathogenicity, and TOR-related autophagy homeostasis in Pyricularia oryzae. Taken together, our results suggested that PoElp3 acts through the tRNA-mediated translational efficiency to regulate asexual development, pathogenicity, sphingolipid metabolism, and autophagy in the rice blast fungus.

Key words: Elongator , tRNA , proteomics , translational efficiency , sphingolipid C9-methyltransferase , rice blast fungus

Yuanhao Liu, Ting Sun, Yuyong Li, Jianqiang Huang, Xianjun Wang, Huimin Bai, Jiayi Hu, Zifan Zhang, Shuai Wang, Dongmei Zhang, Xiuxiu Li, Zonghua Wang, Huakun Zheng, Guifang Lin. 蛋白质组分析揭示了稻瘟病菌延伸复合物蛋白PoElp3通过调节tRNA介导的翻译效率影响发育、致病性和自噬的机制[J]. Journal of Integrative Agriculture, 2025, 24(4): 1515-1528.

Yuanhao Liu, Ting Sun, Yuyong Li, Jianqiang Huang, Xianjun Wang, Huimin Bai, Jiayi Hu, Zifan Zhang, Shuai Wang, Dongmei Zhang, Xiuxiu Li, Zonghua Wang, Huakun Zheng, Guifang Lin. Proteomic analysis revealed the function of PoElp3 in development, pathogenicity, and autophagy through the tRNA-mediated translation efficiency in the rice blast fungus[J]. Journal of Integrative Agriculture, 2025, 24(4): 1515-1528.

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