Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia, United States of America.
Department of Cellular Biology, University of Georgia, Athens, Georgia, United States of America.
PLoS Pathog. 2021 Mar 15;17(3):e1009399. doi: 10.1371/journal.ppat.1009399. eCollection 2021 Mar.
Trypanosoma cruzi is a protist parasite and the causative agent of American trypanosomiasis or Chagas disease. The parasite life cycle in its mammalian host includes an intracellular stage, and glycosylated proteins play a key role in host-parasite interaction facilitating adhesion, invasion and immune evasion. Here, we report that a Golgi-localized Mn2+-Ca2+/H+ exchanger of T. cruzi (TcGDT1) is required for efficient protein glycosylation, host cell invasion, and intracellular replication. The Golgi localization was determined by immunofluorescence and electron microscopy assays. TcGDT1 was able to complement the growth defect of Saccharomyces cerevisiae null mutants of its ortholog ScGDT1 but ablation of TcGDT1 by CRISPR/Cas9 did not affect the growth of the insect stage of the parasite. The defect in protein glycosylation was rescued by Mn2+ supplementation to the growth medium, underscoring the importance of this transition metal for Golgi glycosylation of proteins.
克氏锥虫是一种原生动物寄生虫，也是美洲锥虫病或恰加斯病的病原体。该寄生虫在哺乳动物宿主体内的生命周期包括一个细胞内阶段，糖基化蛋白在促进黏附、入侵和免疫逃逸的宿主-寄生虫相互作用中发挥关键作用。在这里，我们报告称，克氏锥虫的高尔基体定位的 Mn2+-Ca2+/H+ 交换蛋白（TcGDT1）对于有效的蛋白糖基化、宿主细胞入侵和细胞内复制是必需的。高尔基体定位通过免疫荧光和电子显微镜检测来确定。TcGDT1 能够补充其同源物 ScGDT1 的酿酒酵母 null 突变体的生长缺陷，但通过 CRISPR/Cas9 敲除 TcGDT1 并不影响寄生虫的昆虫阶段的生长。通过向生长培养基中补充 Mn2+可以挽救蛋白糖基化缺陷，这突出了这种过渡金属对高尔基体糖基化蛋白的重要性。
