Genome-Wide Bimolecular Fluorescence Complementation-Based Proteomic Analysis of Toxoplasma gondii ROP18's Human Interactome Shows Its Key Role in Regulation of Cell Immunity and Apoptosis.
Genome-Wide Bimolecular Fluorescence Complementation-Based Proteomic Analysis of Toxoplasma gondii ROP18's Human Interactome Shows Its Key Role in Regulation of Cell Immunity and Apoptosis.
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基于全基因组双分子荧光互补的弓形虫 ROP18 人类相互作用组的蛋白质组学分析显示其在细胞免疫和细胞凋亡调节中的关键作用。
DOI：
10.3389/fimmu.2018.00061
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发表时间：
2018
影响因子：
7.3
通讯作者：
Peng HJ
中科院分区：
文献类型：
--
作者：
Xia J;Kong L;Zhou LJ;Wu SZ;Yao LJ;He C;He CY;Peng HJ
弓形虫棒状蛋白ROP18(TgROP18)是弓形虫侵袭过程中分泌到宿主细胞中的一个关键毒力因子，它通过与宿主靶点相互作用来调节宿主细胞的反应。然而，只发现了几个TgROP18目标。在这项研究中，我们应用双分子荧光互补(BIFC)的高通量蛋白质-蛋白质相互作用(PPI)技术在人类细胞中筛选出了I型ROP18株(ROP18I)和II型株ROP18(ROP18II)的靶标。在18,000多个人类蛋白质库中，分别有492个和141个蛋白质被鉴定为ROP18I和ROP18II的靶标。基因本体论、检索相互作用基因/蛋白质的搜索工具PPI网络和独创性途径分析表明，这些蛋白质中的大多数与免疫反应和细胞凋亡有关。这表明TgROP18在调控宿主免疫和细胞凋亡中起着关键作用，这可能有助于寄生虫的免疫逃逸和成功的寄生。在鉴定的蛋白质中，免疫相关蛋白N-myc和STAT相互作用蛋白、IL20RB、IL21、泛素C和波形蛋白以及凋亡相关蛋白P2RX1通过敏化发射-荧光共振能量转移(SE-FRET)和免疫共沉淀进一步证实为ROP18I靶点。我们的研究极大地改善了目前对TgROP18人类靶标的有限了解，并为研究寄生虫效应器在人类细胞中的功能提供了一个新的工具。
Toxoplasma gondii rhoptry protein ROP18 (TgROP18) is a key virulence factor secreted into the host cell during invasion, where it modulates the host cell response by interacting with its host targets. However, only a few TgROP18 targets have been identified. In this study, we applied a high-throughput protein–protein interaction (PPI) screening in human cells using bimolecular fluorescence complementation (BiFC) to identify the targets of Type I strain ROP18 (ROP18I) and Type II strain ROP18 (ROP18II). From a pool of more than 18,000 human proteins, 492 and 141 proteins were identified as the targets of ROP18I and ROP18II, respectively. Gene ontology, search tool for the retrieval of interacting genes/proteins PPI network, and Ingenuity pathway analyses revealed that the majority of these proteins were associated with immune response and apoptosis. This indicates a key role of TgROP18 in manipulating host’s immunity and cell apoptosis, which might contribute to the immune escape and successful parasitism of the parasite. Among the proteins identified, the immunity-related proteins N-myc and STAT interactor, IL20RB, IL21, ubiquitin C, and vimentin and the apoptosis-related protein P2RX1 were further verified as ROP18I targets by sensitized emission-fluorescence resonance energy transfer (SE-FRET) and co-immunoprecipitation. Our study substantially contributes to the current limited knowledge on human targets of TgROP18 and provides a novel tool to investigate the function of parasite effectors in human cells.
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影响因子：
3.1
作者：
HAQUE, S;KHAN, I;KASPER, L
通讯作者：
KASPER, L
影响因子：
3.1
作者：
Howe, DK;Summers, BC;Sibley, LD
通讯作者：
Sibley, LD
影响因子：
3.4
作者：
Howell, JM;Winstone, TL;Turner, RJ
通讯作者：
Turner, RJ
DOI：
10.1038/nrg2918
发表时间：
2011-01
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
5.6
作者：
Citovsky, Vitaly;Lee, Lan-Ying;Tzfira, Tzvi
通讯作者：
Tzfira, Tzvi