Wang Yubao, Weiss Louis M, Orlofsky Amos
Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Biol Chem. 2009 Jan 16;284(3):1694-701. doi: 10.1074/jbc.M807890200. Epub 2008 Nov 21.
Autophagy has been shown to contribute to defense against intracellular bacteria and parasites. In comparison, the ability of such pathogens to manipulate host cell autophagy to their advantage has not been examined. Here we present evidence that infection by Toxoplasma gondii, an intracellular protozoan parasite, induces host cell autophagy in both HeLa cells and primary fibroblasts, via a mechanism dependent on host Atg5 but independent of host mammalian target of rapamycin suppression. Infection led to the conversion of LC3 to the autophagosome-associated form LC3-II, to the accumulation of LC3-containing vesicles near the parasitophorous vacuole, and to the relocalization toward the vacuole of structures labeled by the phosphatidylinositol 3-phosphate indicator YFP-2xFYVE. The autophagy regulator beclin 1 was concentrated in the vicinity of the parasitophorous vacuole in infected cells. Inhibitor studies indicated that parasite-induced autophagy is dependent on calcium signaling and on abscisic acid. At physiologically relevant amino acid levels, parasite growth became defective in Atg5-deficient cells, indicating a role for host cell autophagy in parasite recovery of host cell nutrients. A flow cytometric analysis of cell size as a function of parasite content revealed that autophagy-dependent parasite growth correlates with autophagy-dependent consumption of host cell mass that is dependent on parasite progression. These findings indicate a new role for autophagy as a pathway by which parasites may effectively compete with the host cell for limiting anabolic resources.
自噬已被证明有助于抵御细胞内细菌和寄生虫。相比之下,此类病原体利用宿主细胞自噬为自身谋利的能力尚未得到研究。在此,我们提供证据表明,细胞内原生动物寄生虫刚地弓形虫的感染通过一种依赖宿主Atg5但不依赖宿主雷帕霉素靶蛋白抑制的机制,在HeLa细胞和原代成纤维细胞中诱导宿主细胞自噬。感染导致LC3转化为与自噬体相关的形式LC3-II,导致含LC3的囊泡在寄生泡附近积累,并导致由磷脂酰肌醇3-磷酸指示剂YFP-2xFYVE标记的结构重新定位到泡附近。自噬调节因子贝clin 1在感染细胞的寄生泡附近聚集。抑制剂研究表明,寄生虫诱导的自噬依赖于钙信号和脱落酸。在生理相关的氨基酸水平下,寄生虫在Atg5缺陷细胞中的生长出现缺陷,表明宿主细胞自噬在寄生虫获取宿主细胞营养方面发挥作用。作为寄生虫含量函数的细胞大小的流式细胞术分析表明,自噬依赖性寄生虫生长与依赖于寄生虫进展的宿主细胞质量的自噬依赖性消耗相关。这些发现表明自噬具有一种新作用,即寄生虫可通过该途径与宿主细胞有效竞争有限的合成代谢资源。