Proteomics Reveals AP-2 Complex Depletion Suppressing Listeria monocytogenes Intracellular Replication

IF 3.9 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Proteomics Pub Date : 2025-08-23 DOI:10.1002/pmic.70034

Zhangfu Li, Haiying Ran, Xiangyu Tang, Xiao Liu, Xiaoyuan Wan, Pei Xiong, Zhe Gan, Xu Liu, Liting Wang, Jiangbei Yuan

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引用次数: 0

Abstract

Listeria monocytogenes represents a significant zoonotic pathogen that completes its infectious cycle by invading intestinal epithelial cells, breaching mucosal barriers, and disseminating to target organs such as the liver and spleen. During this process, the innate immune system, particularly macrophages and dendritic cells, plays a pivotal role in pathogen clearance. In this study, we established an in vitro infection model utilizing Raw264.7 macrophages, DC2.4 dendritic cells, HeLa, and Caco-2 epithelial cells. We demonstrated L. monocytogenes infection significantly upregulates the expression of Ap2s1, a key subunit of the AP-2 adaptor complex, in host cells. Subsequent proteomic analysis revealed that two additional AP-2 complex subunits, Ap2m1 and Ap2a2, functionally cooperate with Ap2s1 during infection. Genetic knockout experiments confirmed that specific silencing of any of these three subunits, Ap2m1, Ap2a2, or Ap2s1, markedly inhibited intracellular bacterial proliferation. These findings establish the AP-2 complex as a promising molecular target for developing novel therapeutic strategies against L. monocytogenes infections.

Summary

This study unveils a critical role of the AP-2 adaptor complex in L. monocytogenes intracellular replication, identifying three subunits, Ap2s1, Ap2m1, and Ap2a2, as key host factors exploited by the pathogen.
Using proteomics and CRISPR/Cas9-mediated knockout models, we demonstrate that L. monocytogenes infection upregulates Ap2s1 expression, and depletion of AP-2 subunit significantly impairs bacterial proliferation.
Our findings reveal that AP-2 facilitates post-phagosomal cytosolic replication, independent of bacterial cell-to-cell spread.
This study demonstrates that the AP-2 complex may represent a host factor exploited by L. monocytogenes to promote intracellular replication, offering a novel host-directed therapeutic target to combat antibiotic-resistant strains.

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蛋白质组学揭示AP-2复合物耗竭抑制单核增生李斯特菌胞内复制。

单核增生李斯特菌是一种重要的人畜共患病原体，它通过侵入肠上皮细胞，突破粘膜屏障，并传播到靶器官如肝脏和脾脏来完成其感染周期。在这一过程中，先天免疫系统，特别是巨噬细胞和树突状细胞，在病原体清除中起着关键作用。在本研究中，我们利用Raw264.7巨噬细胞、DC2.4树突状细胞、HeLa和Caco-2上皮细胞建立体外感染模型。我们证明单核增生乳杆菌感染显著上调宿主细胞中AP-2接头复合体的关键亚基Ap2s1的表达。随后的蛋白质组学分析显示，在感染过程中，另外两个AP-2复合物亚基Ap2m1和Ap2a2在功能上与Ap2s1合作。基因敲除实验证实，特异性沉默Ap2m1、Ap2a2或Ap2s1这三个亚基中的任何一个都能显著抑制细胞内细菌的增殖。这些发现确立了AP-2复合体作为开发抗单核细胞增生乳杆菌感染新治疗策略的有希望的分子靶点。摘要：本研究揭示了AP-2接头复合物在单核增生李斯特菌细胞内复制中的关键作用，确定了Ap2s1、Ap2m1和Ap2a2三个亚基是病原菌利用的关键宿主因子。利用蛋白质组学和CRISPR/ cas9介导的敲除模型，我们证明单核增生乳杆菌感染上调Ap2s1的表达，AP-2亚基的缺失显著损害细菌的增殖。我们的研究结果表明，AP-2促进吞噬体后细胞质复制，独立于细菌细胞间扩散。本研究表明，AP-2复合物可能是单核增生乳杆菌促进细胞内复制的宿主因子，为对抗耐药菌株提供了一种新的宿主导向治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Proteomics 生物-生化研究方法

CiteScore

6.30

自引率

5.90%

发文量

193

审稿时长

3 months

期刊介绍： PROTEOMICS is the premier international source for information on all aspects of applications and technologies, including software, in proteomics and other "omics". The journal includes but is not limited to proteomics, genomics, transcriptomics, metabolomics and lipidomics, and systems biology approaches. Papers describing novel applications of proteomics and integration of multi-omics data and approaches are especially welcome.