Zuccala Elizabeth S, Satchwell Timothy J, Angrisano Fiona, Tan Yan Hong, Wilson Marieangela C, Heesom Kate J, Baum Jake
Division of Infection and Immunity, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
Department of Medical Biology, University of Melbourne, Parkville, Victoria, Australia.
Sci Rep. 2016 Feb 2;6:19766. doi: 10.1038/srep19766.
The invasive blood-stage malaria parasite - the merozoite - induces rapid morphological changes to the target erythrocyte during entry. However, evidence for active molecular changes in the host cell that accompany merozoite invasion is lacking. Here, we use invasion inhibition assays, erythrocyte resealing and high-definition imaging to explore red cell responses during invasion. We show that although merozoite entry does not involve erythrocyte actin reorganisation, it does require ATP to complete the process. Towards dissecting the ATP requirement, we present an in depth quantitative phospho-proteomic analysis of the erythrocyte during each stage of invasion. Specifically, we demonstrate extensive increased phosphorylation of erythrocyte proteins on merozoite attachment, including modification of the cytoskeletal proteins beta-spectrin and PIEZO1. The association with merozoite contact but not active entry demonstrates that parasite-dependent phosphorylation is mediated by host-cell kinase activity. This provides the first evidence that the erythrocyte is stimulated to respond to early invasion events through molecular changes in its membrane architecture.
侵入性血液阶段疟原虫——裂殖子——在进入过程中会使靶红细胞迅速发生形态变化。然而,缺乏关于裂殖子入侵时宿主细胞中伴随的主动分子变化的证据。在这里,我们使用入侵抑制试验、红细胞重封和高清成像来探索入侵过程中的红细胞反应。我们发现,虽然裂殖子进入并不涉及红细胞肌动蛋白重组,但确实需要ATP来完成这一过程。为了剖析对ATP的需求,我们对入侵各阶段的红细胞进行了深入的定量磷酸化蛋白质组分析。具体而言,我们证明了裂殖子附着时红细胞蛋白的磷酸化广泛增加,包括细胞骨架蛋白β-血影蛋白和PIEZO1的修饰。与裂殖子接触而非主动进入的关联表明,寄生虫依赖性磷酸化是由宿主细胞激酶活性介导的。这首次证明红细胞通过其膜结构的分子变化被刺激对早期入侵事件做出反应。