Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (IPN), Ciudad de México, Mexico.
Laboratorio de Bioinformática y Simulación Molecular, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Sinaloa, Sinaloa, Mexico.
Front Cell Infect Microbiol. 2024 Sep 27;14:1467440. doi: 10.3389/fcimb.2024.1467440. eCollection 2024.
In humans and , the functional convergence of the endosomal sorting complex required for transport (ESCRT) machinery that is in charge of selecting ubiquitinated proteins for sorting into multivesicular bodies, and the retromer, that is the complex responsible for protein recycling to the plasma membrane and Golgi apparatus. ESCRT and retromer complexes are codependent for protein sorting recycling, degradation, and secretion. In this article, we studied the EhVps35 C isoform (referred to as EhVps35), that is the central member of the retromer, and its relation with the ESCRT machinery during sorting and protein recycling events and their involvement virulence. Our findings revealed that EhVps35 interacts with at least 300 proteins that participate in multiple cellular processes. Laser confocal and transmission electronic microscopy images, as well as secretion assays, revealed that EhVps35 is secreted in vesicles together with EhVps23 and EhADH (both ESCRT machinery proteins). In addition, immunoprecipitation, immunofluorescence, and molecular docking assays revealed the relationship among EhVps35 and other ESCRT machinery proteins. Red blood cell stimulus increased EhVps35 secretion, and the knockdown of the gene in trophozoites reduced their capacity to migrate and invade tissues. This also impacts the cellular localization of ubiquitin, EhVps23 (ESCRT-I), and EhVps32 (ESCRT-III) proteins, strongly suggesting their functional relationship. Our results, taken together, give evidence that EhVps35 is a key factor in virulence mechanisms and that it, together with the ESCRT machinery components and other regulatory proteins, is involved in vesicle trafficking, secretion, migration, and cell proliferation.
在人类和中，负责选择泛素化蛋白进行分类到多泡体的内体分选复合物所需的功能（ESCRT）机制，以及负责将蛋白质回收至质膜和高尔基体的复合物（retromer），这两个复合物在蛋白质分类回收、降解和分泌方面是相互依赖的。在本文中，我们研究了 EhVps35 C 异构体（称为 EhVps35），它是 retromer 的核心成员，以及其在分选和蛋白质回收事件中的 ESCRT 机制之间的关系及其参与毒力。我们的研究结果表明，EhVps35 与至少 300 种参与多种细胞过程的蛋白质相互作用。激光共聚焦和透射电子显微镜图像以及分泌测定显示，EhVps35 与 EhVps23 和 EhADH（ESCRT 机制蛋白）一起分泌到囊泡中。此外，免疫沉淀、免疫荧光和分子对接测定揭示了 EhVps35 与其他 ESCRT 机制蛋白之间的关系。红细胞刺激增加了 EhVps35 的分泌，而滋养体中基因的敲低降低了它们迁移和侵袭组织的能力。这也影响了泛素、EhVps23（ESCRT-I）和 EhVps32（ESCRT-III）蛋白的细胞定位，强烈表明它们的功能关系。我们的研究结果表明，EhVps35 是 毒力机制的关键因素，它与 ESCRT 机制成分和其他调节蛋白一起，参与囊泡运输、分泌、迁移和细胞增殖。
