Pernas Lena, Adomako-Ankomah Yaw, Shastri Anjali J, Ewald Sarah E, Treeck Moritz, Boyle Jon P, Boothroyd John C
Department of Microbiology & Immunology, Stanford University School of Medicine, Stanford, California, United States of America.
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
PLoS Biol. 2014 Apr 29;12(4):e1001845. doi: 10.1371/journal.pbio.1001845. eCollection 2014 Apr.
Recent information has revealed the functional diversity and importance of mitochondria in many cellular processes including orchestrating the innate immune response. Intriguingly, several infectious agents, such as Toxoplasma, Legionella, and Chlamydia, have been reported to grow within vacuoles surrounded by host mitochondria. Although many hypotheses have been proposed for the existence of host mitochondrial association (HMA), the causes and biological consequences of HMA have remained unanswered. Here we show that HMA is present in type I and III strains of Toxoplasma but missing in type II strains, both in vitro and in vivo. Analysis of F1 progeny from a type II×III cross revealed that HMA is a Mendelian trait that we could map. We use bioinformatics to select potential candidates and experimentally identify the polymorphic parasite protein involved, mitochondrial association factor 1 (MAF1). We show that introducing the type I (HMA+) MAF1 allele into type II (HMA-) parasites results in conversion to HMA+ and deletion of MAF1 in type I parasites results in a loss of HMA. We observe that the loss and gain of HMA are associated with alterations in the transcription of host cell immune genes and the in vivo cytokine response during murine infection. Lastly, we use exogenous expression of MAF1 to show that it binds host mitochondria and thus MAF1 is the parasite protein directly responsible for HMA. Our findings suggest that association with host mitochondria may represent a novel means by which Toxoplasma tachyzoites manipulate the host. The existence of naturally occurring HMA+ and HMA- strains of Toxoplasma, Legionella, and Chlamydia indicates the existence of evolutionary niches where HMA is either advantageous or disadvantageous, likely reflecting tradeoffs in metabolism, immune regulation, and other functions of mitochondria.
近期的信息揭示了线粒体在包括协调先天性免疫反应在内的许多细胞过程中的功能多样性和重要性。有趣的是,据报道,几种感染因子,如弓形虫、嗜肺军团菌和衣原体,能在被宿主线粒体包围的液泡内生长。尽管对于宿主线粒体关联(HMA)的存在提出了许多假说,但HMA的成因和生物学后果仍未得到解答。在这里,我们表明,无论是在体外还是体内,HMA存在于弓形虫的I型和III型菌株中,而在II型菌株中缺失。对II型×III型杂交的F1后代的分析表明,HMA是一种我们可以定位的孟德尔性状。我们使用生物信息学来选择潜在的候选基因,并通过实验鉴定出所涉及的多态性寄生虫蛋白——线粒体关联因子1(MAF1)。我们表明,将I型(HMA+)MAF1等位基因引入II型(HMA-)寄生虫会导致其转变为HMA+,而在I型寄生虫中删除MAF1会导致HMA丧失。我们观察到,HMA的丧失和获得与宿主细胞免疫基因转录的改变以及小鼠感染期间的体内细胞因子反应有关。最后,我们通过MAF1的外源表达表明它与宿主线粒体结合,因此MAF1是直接导致HMA的寄生虫蛋白。我们的研究结果表明,与宿主线粒体的关联可能代表了速殖子弓形虫操纵宿主 的一种新方式。弓形虫、嗜肺军团菌和衣原体自然存在的HMA+和HMA-菌株的存在表明存在HMA有利或不利的进化生态位,这可能反映了线粒体在代谢、免疫调节和其他功能方面的权衡。