Use of RNA interference in Drosophila S2 cells to identify host pathways controlling compartmentalization of an intracellular pathogen
Use of RNA interference in Drosophila S2 cells to identify host pathways controlling compartmentalization of an intracellular pathogen
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DOI：
10.1073/pnas.0506461102
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发表时间：
2005-09-20
影响因子：
11.1
通讯作者：
Portnoy, DA
中科院分区：
文献类型：
--
作者：
Cheng, LW;Viala, JPM;Portnoy, DA
在果蝇S2细胞中进行了三种全基因组RNA干扰筛选，以剖析宿主过程对单核细胞增多性李斯特菌进入、空泡逃逸和细胞内生长的贡献。在已鉴定的116个基因中，已确定了几个先前未被认为在李斯特菌致病过程中起作用的宿主途径：影响空泡进出多囊体内的必需成孔毒素李斯特溶素O的敲除和影响丝氨酸棕榈酰转氨酶(神经酰胺和神经鞘糖脂生物合成的关键酶)亚单位的敲除，增强了在宿主细胞胞浆中表达的李斯特溶素O的毒性，导致缺乏适当的毒素活性区隔和宿主细胞死亡。利用果蝇S2细胞进行全基因组RNA干扰筛选被证明是剖析宿主-病原体相互作用的一种强有力的方法。
Three genome-wide RNA interference screens were performed in Drosophila S2 cells to dissect the contribution of host processes to Listeria monocytogenes entry, vacuolar escape, and intracellular growth. Among the 116 genes identified, several host pathways previously unrecognized as playing a role in listerial pathogenesis were identified: knockdowns affecting vacuolar trafficking to and from the multivesicular body bypassed the requirement for the essential pore-forming toxin listeriolysin O in mediating escape from phagocytic vacuoles and knockdowns affecting either subunit of serine palm itoyltra nsferase, a key enzyme in ceramide and sphingolipid biosynthesis, enhanced the toxicity of listeriolysin O expressed in the host cell cytosol, leading to lack of appropriate toxin activity compartmentalization and host cell death. Genome-wide RNA interference screens using Drosophila S2 cells proved to be a powerful approach to dissect host-pathogen interactions.