Giuseppe
Emeritus
[Source: PLoS ONE, full text: (LINK). Abstract, edited.]
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Influenza Virus A/Beijing/501/2009(H1N1) NS1 Interacts with β-Tubulin and Induces Disruption of the Microtubule Network and Apoptosis on A549 Cells
Xueqing Han<SUP>1</SUP><SUP>#</SUP>, Zhihui Li<SUP>1</SUP><SUP>#</SUP>, Hongjun Chen<SUP>1</SUP>, Huiyu Wang<SUP>1</SUP>, Lin Mei<SUP>1</SUP>, Shaoqiang Wu<SUP>1</SUP>, Tianyi Zhang<SUP>3</SUP>, Bohua Liu<SUP>2</SUP><SUP>*</SUP>, Xiangmei Lin<SUP>1</SUP><SUP>*</SUP>
<SUP></SUP>
1 Chinese Academy of Inspection and Quarantine, Beijing, China, 2 State Key Laboratory of Pathogens and Bio-security, Beijing Institute of Microbiology and Epidemiology, Beijing, China, 3 College of Veterinary Medicine, China Agricultural University, Bejing, China
Abstract
NS1 of influenza A virus is a key multifunctional protein that plays various roles in regulating viral replication mechanisms, host innate/adaptive immune responses, and cellular signalling pathways. These functions rely on its ability to participate in a multitude of protein-protein and protein-RNA interactions. To gain further insight into the role of NS1, a tandem affinity purification (TAP) method was utilized to find unknown interaction partner of NS1. The protein complexes of NS1 and its interacting partner were purified from A549 cell using TAP-tagged NS1 as bait, and co-purified cellular factors were identified by mass spectrometry (MS). We identified cellular β-tubulin as a novel interaction partner of NS1. The RNA-binding domain of NS1 interacts with β-tubulin through its RNA-binding domain, as judged by a glutathione S-transferase (GST) pull-down assay with the GST-fused functional domains of NS1. Immunofluorescence analysis further revealed that NS1 with β-tubulin co-localized in the nucleus. In addition, the disruption of the microtubule network and apoptosis were also observed on NS1-transfected A549 cells. Our findings suggest that influenza A virus may utilize its NS1 protein to interact with cellular β-tubulin to further disrupt normal cell division and induce apoptosis. Future work will illustrate whether this interaction is uniquely specific to the 2009 pandemic H1N1 virus.
Citation: Han X, Li Z, Chen H, Wang H, Mei L, et al. (2012) Influenza Virus A/Beijing/501/2009(H1N1) NS1 Interacts with β-Tubulin and Induces Disruption of the Microtubule Network and Apoptosis on A549 Cells. PLoS ONE 7(11): e48340. doi:10.1371/journal.pone.0048340
Editor: Michael C. W. Chan, Centre of Influenza Research, The University of Hong Kong, Hong Kong
Received: June 6, 2012; Accepted: September 24, 2012; Published: November 6, 2012
Copyright: ? 2012 Han et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the National Basic Research Program of China (973 Program) (Grant No. 2011CB504704), http://www.973.gov.cn/AreaAppl.aspx, and the Fundamental Research Fund of the Chinese Academy of Inspection and Quarantine (Grant No. 2011JK004), http://www.caiq.gov.cn/jky/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
* E-mail: xiangmei8lin@yahoo.com.cn (XL); bhliuamms@yahoo.com.cn (BL)
# These authors contributed equally to this work.
-Xueqing Han<SUP>1</SUP><SUP>#</SUP>, Zhihui Li<SUP>1</SUP><SUP>#</SUP>, Hongjun Chen<SUP>1</SUP>, Huiyu Wang<SUP>1</SUP>, Lin Mei<SUP>1</SUP>, Shaoqiang Wu<SUP>1</SUP>, Tianyi Zhang<SUP>3</SUP>, Bohua Liu<SUP>2</SUP><SUP>*</SUP>, Xiangmei Lin<SUP>1</SUP><SUP>*</SUP>
<SUP></SUP>
1 Chinese Academy of Inspection and Quarantine, Beijing, China, 2 State Key Laboratory of Pathogens and Bio-security, Beijing Institute of Microbiology and Epidemiology, Beijing, China, 3 College of Veterinary Medicine, China Agricultural University, Bejing, China
Abstract
NS1 of influenza A virus is a key multifunctional protein that plays various roles in regulating viral replication mechanisms, host innate/adaptive immune responses, and cellular signalling pathways. These functions rely on its ability to participate in a multitude of protein-protein and protein-RNA interactions. To gain further insight into the role of NS1, a tandem affinity purification (TAP) method was utilized to find unknown interaction partner of NS1. The protein complexes of NS1 and its interacting partner were purified from A549 cell using TAP-tagged NS1 as bait, and co-purified cellular factors were identified by mass spectrometry (MS). We identified cellular β-tubulin as a novel interaction partner of NS1. The RNA-binding domain of NS1 interacts with β-tubulin through its RNA-binding domain, as judged by a glutathione S-transferase (GST) pull-down assay with the GST-fused functional domains of NS1. Immunofluorescence analysis further revealed that NS1 with β-tubulin co-localized in the nucleus. In addition, the disruption of the microtubule network and apoptosis were also observed on NS1-transfected A549 cells. Our findings suggest that influenza A virus may utilize its NS1 protein to interact with cellular β-tubulin to further disrupt normal cell division and induce apoptosis. Future work will illustrate whether this interaction is uniquely specific to the 2009 pandemic H1N1 virus.
Citation: Han X, Li Z, Chen H, Wang H, Mei L, et al. (2012) Influenza Virus A/Beijing/501/2009(H1N1) NS1 Interacts with β-Tubulin and Induces Disruption of the Microtubule Network and Apoptosis on A549 Cells. PLoS ONE 7(11): e48340. doi:10.1371/journal.pone.0048340
Editor: Michael C. W. Chan, Centre of Influenza Research, The University of Hong Kong, Hong Kong
Received: June 6, 2012; Accepted: September 24, 2012; Published: November 6, 2012
Copyright: ? 2012 Han et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the National Basic Research Program of China (973 Program) (Grant No. 2011CB504704), http://www.973.gov.cn/AreaAppl.aspx, and the Fundamental Research Fund of the Chinese Academy of Inspection and Quarantine (Grant No. 2011JK004), http://www.caiq.gov.cn/jky/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
* E-mail: xiangmei8lin@yahoo.com.cn (XL); bhliuamms@yahoo.com.cn (BL)
# These authors contributed equally to this work.
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