Screening and Identification of the Host Proteins Interacting with Toxoplasma gondii Rhoptry Protein ROP16.
Screening and Identification of the Host Proteins Interacting with Toxoplasma gondii Rhoptry Protein ROP16.
复制标题
弓形虫棒状体蛋白ROP16相互作用宿主蛋白的筛选与鉴定
DOI：
10.3389/fmicb.2017.02408
复制
发表时间：
2017
影响因子：
5.2
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
Pan M;Zhou Y;Wang Y;Li L;Song Y;Hou L;Zhao J
弓形虫作为一种人畜共患的原生动物寄生虫，开发出复杂的策略来操纵宿主以实现有效的细胞内生存。成功入侵后，弓形虫会出于各种目的将许多效应蛋白注入宿主细胞。 TgROP16（弓形虫棒状体蛋白16）从棒状体分泌到宿主细胞中，可以通过STAT3和STAT6的磷酸化激活宿主STAT（信号转导和转录激活剂）信号通路。然而，目前尚不清楚是否还有其他受TgROP16调节的宿主蛋白。在这项研究中，酵母双杂交 (Y2H) 筛选用于寻找与 TgROP16 相互作用的其他宿主蛋白。表达克隆到猎物载体中的小鼠 cDNA 文库的酵母细胞用于与表达不带信号肽的 ROP16 的酵母交配。从该筛选中鉴定出两种小鼠蛋白 Dnaja1（DnaJ 热休克蛋白家族成员 A1）和 Gabra4（γ-氨基丁酸 A 受体，亚基 α 4）与 ROP16 相互作用。进一步的分析表明，ROP16 的前结构域在介导与这些宿主蛋白的相互作用中发挥着关键作用，而激酶结构域的贡献较小。还通过免疫共沉淀在体内评估了 Dnaja1 和 ROP16 不同部分之间的相互作用。结果表明，ROP16的Predomain是与Dnaja1相互作用的主要区域，这与Y2H的结果一致。基于基因本体分析，预测Dnaja1参与应激反应，而Gabra4参与系统发育过程。与弓形虫ROP16相互作用的新宿主蛋白的发现将有助于我们进一步研究该效应蛋白在弓形虫感染过程中的功能。
Toxoplasma gondii, as a zoonotic protozoan parasite, develops sophisticated strategies to manipulate hosts for efficient intracellular survival. After successful invasion, T. gondii injects many effector proteins into host cells for various purposes. TgROP16 (T. gondii rhoptry protein 16), which is secreted from rhoptries into host cells, can activate the host STAT (signal transducer and activator of transcription) signaling pathway through phosphorylation of STAT3 and STAT6. However, whether there are other host proteins modulated by TgROP16 is currently unknown. In this study, yeast two-hybrid (Y2H) screen was used to look for additional host proteins interacting with TgROP16. Yeast cells expressing a mouse cDNA library cloned into the prey vector were used to mate with yeasts expressing ROP16 without signal peptide. Two mouse proteins, Dnaja1 (DnaJ heat shock protein family member A1) and Gabra4 (gamma-aminobutyric acid A receptor, subunit alpha 4) were identified to interact with ROP16 from this screen. Further analysis suggested that the Predomain of ROP16 played key roles in mediating interactions with these host proteins, whereas the contribution from the Kinase domain was minor. The interactions between Dnaja1 and different parts of ROP16 were also estimated in vivo by co-immunoprecipitation. The results showed that the Predomain of ROP16 was the major region to interact with Dnaja1, which is consistent with the Y2H results. Based on the gene ontology analysis, Dnaja1 is predicted to participate in stress response while Gabra4 is involved in the system development process. The discovery of new host proteins that interact with ROP16 of T. gondii will help us to further investigate the functions of this effector proteins during T. gondii infection.
登录
查看更多内容
戒酒过程中GABRA4表达的下调是由培养的小鼠皮质神经元中的特定microRNA介导的。
影响因子：
3.1
作者：
Bekdash RA;Harrison NL
通讯作者：
Harrison NL
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.
影响因子：
64.8
作者：
Saeij, J. P. J.;Coller, S.;Boothroyd, J. C.
通讯作者：
Boothroyd, J. C.
酵母双杂交系统鉴定与弓形虫 GRA15 (TgGRA15) 相互作用的宿主蛋白
影响因子：
3.2
作者：
Liu Q;Li FC;Elsheikha HM;Sun MM;Zhu XQ
通讯作者：
Zhu XQ
影响因子：
4.4
作者：
Iyer, Sangeetha V.;Chandra, Dave;Homanics, Gregg E.
通讯作者：
Homanics, Gregg E.