A novel high throughput invasion screen identifies host actin regulators required for efficient cell entry by Toxoplasma gondii.
A novel high throughput invasion screen identifies host actin regulators required for efficient cell entry by Toxoplasma gondii.
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DOI：
10.1371/journal.pone.0064693
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Carruthers VB
中科院分区：
文献类型：
--
作者：
Gaji RY;Huynh MH;Carruthers VB
刚地弓形虫严重依赖细胞入侵作为一种生存策略，以在感染过程中逃避免疫清除。尽管人们普遍认为弓形虫的入侵是由寄生虫主导的，而宿主细胞在很大程度上是被动的受害者，但最近的研究表明，宿主成分如微丝和微管确实对入侵有促进作用。因此，为了确定其他宿主因子，我们使用一种基于诱导型荧光素酶的新型入侵检测方法，对一个靶向可成药蛋白质的人类小干扰RNA文库进行了高通量小干扰RNA筛选。对初次筛选中显示入侵能力下降最显著的前100个命中靶点进行了二次筛选确认，揭示了24种可能参与弓形虫进入宿主细胞的蛋白质。有趣的是，其中6个命中靶点似乎通过改变宿主细胞肌动蛋白动力学来影响寄生虫入侵，导致F - 肌动蛋白在细胞周边的沉积增加。这些发现支持了一个新兴的观点，即宿主肌动蛋白动力学对弓形虫入侵很重要，同时还确定了几个可能参与寄生虫入侵的新型宿主因子。
Toxoplasma gondii critically relies on cell invasion as a survival strategy to evade immune clearance during infection. Although it was widely thought that Toxoplasma entry is parasite directed and that the host cell is largely a passive victim, recent studies have suggested that host components such as microfilaments and microtubules indeed contribute to entry. Hence to identify additional host factors, we performed a high-throughput siRNA screen of a human siRNA library targeting druggable proteins using a novel inducible luciferase based invasion assay. The top 100 hits from the primary screen that showed the strongest decreases in invasion were subjected to confirmation by secondary screening, revealing 24 proteins that are potentially involved in Toxoplasma entry into host cells. Interestingly, 6 of the hits appear to affect parasite invasion by modifying host cell actin dynamics, resulting in increased deposition of F-actin at the periphery of the cell. These findings support the emerging notion that host actin dynamics are important for Toxoplasma invasion along with identifying several novel host factors that potentially participate in parasite entry.
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影响因子：
48
作者：
Andenmatten, Nicole;Egarter, Saskia;Jackson, Allison J;Jullien, Nicolas;Herman, Jean-Paul;Meissner, Markus
通讯作者：
Meissner, Markus
影响因子：
3.1
作者：
Carruthers, VB;Håkansson, S;Sibley, LD
通讯作者：
Sibley, LD
DOI：
10.1046/j.1432-1033.2002.03206.x
发表时间：
2002-10-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Ek, P;Pettersson, G;Zetterqvist, Ö
通讯作者：
Zetterqvist, Ö
影响因子：
3.1
作者：
FURTADO, GC;SLOWIK, M;JOINER, KA
通讯作者：
JOINER, KA
影响因子：
11.4
作者：
Helfer, E;Nevalainen, EM;Carlier, MF
通讯作者：
Carlier, MF