Department of Biology, Indiana University, Bloomington, Indiana United States of America.
PLoS Pathog. 2024 Mar 28;20(3):e1011245. doi: 10.1371/journal.ppat.1011245. eCollection 2024 Mar.
The most common intracellular bacterial infection is Wolbachia pipientis, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector-WalE1, which encodes an alpha-synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression of WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates in the host cell cytosol. We next show that WalE1 co-immunoprecipitates with the host protein Past1, although might not directly interact with it, and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO3, a hallmark of endocytosis defects. We also show that flies expressing WalE1 suffer from endocytosis defects in larval nephrocytes. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between Wolbachia and a host protein involved in endocytosis and point to yet another important host cell process impinged upon by Wolbachia's WalE1 effector.
最常见的细胞内细菌感染是沃尔巴克氏体(Wolbachia pipientis),一种操纵宿主生殖并被用于控制昆虫媒介的微生物。沃尔巴克氏体诱导的表型已经研究了几十年,范围从精子-卵子不相容到雄性致死。沃尔巴克氏体如何改变宿主生物学的机制还不太清楚。以前,我们鉴定了第一个沃尔巴克氏体效应物-WalE1,它在 N 端编码一个α-突触核蛋白结构域。纯化的 WalE1 与肌动蛋白沉淀并将其捆绑,当在果蝇中异源表达时,可增加发育中的卵母细胞中的沃尔巴克氏体滴度。在这项工作中,我们首先通过感染果蝇细胞和整个动物的沃尔巴克氏体来鉴定 WalE1 的天然表达。WalE1 在宿主细胞胞质溶胶中呈现为聚集体。我们接下来表明,WalE1 与宿主蛋白 Past1 共免疫沉淀,尽管可能没有直接相互作用,但 WalE1 操纵宿主内吞作用。表达 WalE1 的酵母显示 FM4-64 染料摄取缺陷,而过表达 WalE1 或 Past1 突变的果蝇对 AgNO3 敏感,这是内吞作用缺陷的标志。我们还表明,表达 WalE1 的果蝇在幼虫肾细胞中遭受内吞作用缺陷。最后,我们还表明,Past1 缺失的果蝇总体上和在晚期卵囊中含有更多的沃尔巴克氏体。我们的结果鉴定了沃尔巴克氏体与参与内吞作用的宿主蛋白之间的相互作用,并指出沃尔巴克氏体的 WalE1 效应物又影响了一个重要的宿主细胞过程。