Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system.
Identification of host proteins interacting with Toxoplasma gondii GRA15 (TgGRA15) by yeast two-hybrid system.
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酵母双杂交系统鉴定与弓形虫 GRA15 (TgGRA15) 相互作用的宿主蛋白
DOI：
10.1186/s13071-016-1943-1
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发表时间：
2017-01-03
影响因子：
3.2
通讯作者：
Zhu XQ
中科院分区：
文献类型：
--
作者：
Liu Q;Li FC;Elsheikha HM;Sun MM;Zhu XQ
背景：刚地弓形虫是一种专性胞内寄生的原虫，具有显著的利用宿主细胞机制以维持其胞内生存的能力。该寄生虫可以通过分泌位于棒状体和致密颗粒细胞器中的多态性效应蛋白来调节其宿主的信号传导途径。T.弓形虫II型特异性致密颗粒蛋白15，TgGRA 15，其激活NF-κB通路。本研究旨在鉴定TgGRA 15的宿主相互作用伴侣蛋白。方法：以TgGRA 15为诱饵，从酵母双杂交小鼠cDNA文库中筛选TgGRA 15。将TgGRA 15（PRU菌株，II型）克隆到pGBKT 7载体中并在Y2 HGold酵母菌株中表达。结果：筛选出小鼠Luzp 1和AW 209491作为与TgGRA 15相互作用的宿主结合蛋白。Luzp 1包含三个核定位信号，并参与调节宿主非编码RNA基因的一个子集。结论：这些研究结果揭示，第一次，新的宿主细胞蛋白与TgGRA 15相互作用。这些细胞靶点的鉴定和对它们在宿主-病原体相互作用中的作用的理解可能为针对T.弓形虫感染
Background:Toxoplasma gondii, an obligate intracellular protozoan parasite, possesses the remarkable ability to co-opt host cell machinery in order to maintain its intracellular survival. This parasite can modulate signaling pathways of its host through the secretion of polymorphic effector proteins localized in the rhoptry and dense granule organelles. One of such effectors is T. gondii type II-specific dense granule protein 15, TgGRA15, which activates NF-κB pathway. The aim of the present study was to identify the host interaction partner proteins of TgGRA15.Methods:We screened a yeast two-hybrid mouse cDNA library using TgGRA15 as the bait. TgGRA15 (PRU strain, Type II) was cloned into the pGBKT7 vector and expressed in the Y2HGold yeast strain. Then, the bait protein expression was validated by western blotting analysis, followed by auto-activation and toxicity tests in comparison with control (Y2HGold yeast strain transformed with empty pGBKT7 vector).Results:This screening led to the identification of mouse Luzp1 and AW209491 as host binding proteins that interact with TgGRA15. Luzp1 contains three nuclear localizing signals and is involved in regulating a subset of host non-coding RNA genes.Conclusions:These findings reveal, for the first time, new host cell proteins interacting with TgGRA15. The identification of these cellular targets and the understanding of their contribution to the host-pathogen interaction may serve as the foundation for novel therapeutic and prevention strategies against T. gondii infection.
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影响因子：
6.4
作者：
Gov L;Karimzadeh A;Ueno N;Lodoen MB
通讯作者：
Lodoen MB
影响因子：
3.2
作者：
Liu Q;Wang ZD;Huang SY;Zhu XQ
通讯作者：
Zhu XQ
影响因子：
3.1
作者：
Jensen, Kirk D. C.;Hu, Kenneth;Saeij, Jeroen P. J.
通讯作者：
Saeij, Jeroen P. J.
DOI：
10.1111/tra.12049
发表时间：
2013-05
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Hsiao CH;Luisa Hiller N;Haldar K;Knoll LJ
通讯作者：
Knoll LJ
影响因子：
3.2
作者：
Chen J;Li ZY;Zhou DH;Liu GH;Zhu XQ
通讯作者：
Zhu XQ