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Dengue virus infection induces formation of G3BP1 granules in human lung epithelial cells

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Abstract

Cells reprogram ongoing translation in response to viral infection, resulting in formation of stress granules (SGs), while viruses have evolved a variety of strategies to antagonize the host SG response. Previous literature reported that in BHK-1 cells, infection with dengue virus (DENV) interfered with the SG formation. In the current study, we further investigated SG formation in human epithelial A549 cells by detecting subcellular localization of two SG hallmarks, TIA-1 and G3BP1. In response to DENV type 2 (DENV2) and type 3 (DENV3) infection, G3BP1, but not TIA-1, was recruited into cytoplasmic granules in some cells, and viral protein synthesis was significantly impaired in the G3BP1-granule-containing cells. Knockdown of G3BP1 significantly rescued the dsRNA-mediated suppression of DENV2 replication. Furthermore, our data showed that the phosphorylation of protein kinase regulated by dsRNA (PKR) and eIF2α, as well as accumulation of dsRNA, mainly occurred at the late stage of viral infection. This work revealed that in DENV-infected A549 cells, G3BP1 granules were assembled independently of TIA-1 and had a negative impact on viral replication. This extends our understanding of the antagonistic relationship between the SG response and dengue virus infection.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (81171576, 81371794), and Guangdong Natural Science Foundation (S2013010016454, 2014A030311007). The NS1 monoclonal Ab was generously provided by Prof. Trai-Ming Yeh (National Cheng Kung University, Taiwan).

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    Authors and Affiliations

    1. Department of Immunology, Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, China

      Jun Xia, Xiaoyan Chen, Feng Xu, Yi Wang, Yuye Li, Junfang He & Ping Zhang

    2. Key Laboratory of Tropical Diseases Control, Ministry of Education, Guangzhou, 510080, China

      Jun Xia, Xiaoyan Chen, Feng Xu, Yi Wang, Yuye Li, Junfang He & Ping Zhang

    3. Guangdong Inspection and Quarantine Technology Center, Guangzhou, 510080, China

      Yongxia Shi

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    1. Jun Xia
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    Correspondence to Junfang He or Ping Zhang.

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    J. Xia and X. Chen contributed equally to this work.

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    705_2015_2578_MOESM1_ESM.tif

    Figure S1. Time course of subcellular localization of TIA-1 and G3BP1 in mock-infected cells. A549 cells were mock infected, harvested at the indicated times, and processed for IFM using anti-TIA-1 (green) and anti-NS1(red) (A) or anti-G3BP1 (red) and anti-NS-1 (green) (B). The images are representative of three independent experiments. Scale bars: 10 μm (TIFF 1201 kb)

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    Xia, J., Chen, X., Xu, F. et al. Dengue virus infection induces formation of G3BP1 granules in human lung epithelial cells. Arch Virol 160, 2991–2999 (2015). https://doi.org/10.1007/s00705-015-2578-9

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