鸡G3BP1在新城疫病毒感染诱导应激颗粒形成过程中的作用

doi:10.11843/j.issn.0366-6964.2017.03.015

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (3): 515-521.doi: 10.11843/j.issn.0366-6964.2017.03.015

鸡G3BP1在新城疫病毒感染诱导应激颗粒形成过程中的作用

张频¹，孙英杰²，郑航³，毛旭明²，仇旭升²，谭磊²，孟春春²，宋翠萍²，廖瑛²，丁铲^2*，马卫明^1*

（1.山东农业大学动物科技学院，泰安 271018；2.中国农业科学院上海兽医研究所，上海 200241；3. 吉林农业大学动物科学技术学院，长春 130118）

收稿日期:2016-10-09 出版日期:2017-03-23 发布日期:2017-03-23
通讯作者: 马卫明，副研究员，Tel：0538-8249222-8205，E-mail:mawm@sdau.edu.cn；丁铲，研究员，Tel：021-34293441，E-mail: shoveldeen@shvri.ac.cn
作者简介:张频（1989-），女，山东菏泽人，硕士生，主要从事动物病毒分子生物学研究，E-mail：794094331@qq.com
基金资助:
国家自然科学基金重点项目（31530074）；国家自然科学基金面上项目（31372421）；国家自然科学基金青年基金（31400144）

The Role of Chicken G3BP1 in the Formation of Stress Granules Induced by Newcastle Disease Virus Infection

ZHANG Pin¹, SUN Ying-jie², ZHENG Hang³, MAO Xu-ming², QIU Xu-sheng², TAN Lei², MENG Chun-chun², SONG Cui-ping², LIAO Ying², DING Chan^2* , MA Wei-ming^1*

(1. College of Animal Science and Technology, Shandong Agricultural University, Tai’an 271018， China; 2. Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China; 3. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China)

Received:2016-10-09 Online:2017-03-23 Published:2017-03-23

摘要/Abstract

摘要：

为掌握鸡细胞中应激颗粒（stress granules，SGs）形成机制，以及鸡GTP酶激活蛋白SH3功能区结合蛋白1（G3BP1）在新城疫病毒（NDV）感染诱导SG形成过程中的作用，用NDV感染HeLa细胞，免疫荧光试验检测内源性G3BP1的定位，外源转染鸡G3BP1和各分段结构域，以不同应激处理之后观察与内源性SG标志物共定位现象，最后转染G3BP1之后感染NDV，以Western blot和TCID₅₀检测病毒复制情况。结果显示：NDV感染能够诱导内源性G3BP1聚集形成SG，将鸡G3BP1分别转染至HeLa、DF-1和CEF后，以不同应激处理之后均能在细胞中观察到SG。将鸡G3BP1不同结构域分别转染HeLa细胞，以氧化应激和NDV分别处理，结果显示G3BP1的RNA结合结构域对聚集至关重要，而NTF2样结构域不能被招募到SGs中，并且抑制原有SGs的形成。最后证实G3BP1过表达能够增强病毒复制，说明SG在NDV复制过程中发挥促进作用。综合以上结果表明：鸡G3BP1在鸡SG形成过程中至关重要，鸡SG能够促进NDV复制，这为进一步研究NDV和禽源细胞互作奠定了基础。

Abstract:

To investigate the mechanism of chicken stress granule (SG) formation and the role of Ras GTPase- activating protein-binding protein 1 (G3BP1) in SG formation and virus infection, immunofluorescence assay was utilized to evaluate G3BP1 accumulation in Newcastle disease virus (NDV) infected HeLa cells. GFP-tagged chicken G3BP1 and its 4 domains were transfected into HeLa cells, respectively, before exposed to various stressors. Finally, G3BP1 was transfected into cells before NDV infection. Western blot and TCID₅₀ test were utilized to evaluate virus replication. The results showed that NDV infection induced endogenous G3BP1 accumulating as SGs. Transfection of exogenous chicken G3BP1 into either HeLa, DF-1 or CEF cells induced SGs in the presence or absence of environmental stress. Then the role of 4 domains of chicken G3BP1 in SG formation was evaluated. The results showed that RNA-binding domain was responsible for SG accumulation. In contrast, NTF2-like domain inhibit SG formation. Finally, it confirmed that G3BP1 overexpression enhanced NDV replication, indicating the pro-viral role of SG in NDV infection. These results demonstrated that chicken G3BP1 plays an important role in SG formation. Chicken SG promotes NDV replication.This study lay the foundation for the interactions between NDV and chicken cells.

中图分类号:

S852.659.5

张频，孙英杰，郑航，毛旭明，仇旭升，谭磊，孟春春，宋翠萍，廖瑛，丁铲，马卫明. 鸡G3BP1在新城疫病毒感染诱导应激颗粒形成过程中的作用[J]. 畜牧兽医学报, 2017, 48(3): 515-521.

ZHANG Pin, SUN Ying-jie, ZHENG Hang, MAO Xu-ming, QIU Xu-sheng, TAN Lei, MENG Chun-chun, SONG Cui-ping, LIAO Ying, DING Chan, MA Wei-ming. The Role of Chicken G3BP1 in the Formation of Stress Granules Induced by Newcastle Disease Virus Infection[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 515-521.

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鸡G3BP1在新城疫病毒感染诱导应激颗粒形成过程中的作用

The Role of Chicken G3BP1 in the Formation of Stress Granules Induced by Newcastle Disease Virus Infection

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