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Proteomic analysis of BmN cell lipid rafts reveals roles in Bombyx mori nucleopolyhedrovirus infection

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Abstract

The mechanism of how Bombyx mori nucleopolyhedrovirus (BmNPV) enters cells is unknown. The primary components of membrane lipid rafts are proteins and cholesterol, and membrane lipid rafts are thought to be an active region for host–viral interactions. However, whether they contribute to the entry of BmNPV into silkworm cells remains unclear. In this study, we explored the membrane protein components of lipid rafts from BmN cells with mass spectrometry (MS). Proteins and cholesterol were investigated after establishing infection with BmNPV in BmN cells. In total, 222 proteins were identified in the lipid rafts, and Gene Ontology (GO) annotation analysis showed that more than 10% of these proteins had binding and catalytic functions. We then identified proteins that potentially interact between lipid rafts and BmNPV virions using the Virus Overlay Protein Blot Assay (VOPBA). A total of 65 proteins were analyzed with MS, and 7 were predicted to be binding proteins involved in BmNPV cellular invasion, including actin, kinesin light chain-like isoform X2, annexin B13, heat-shock protein 90, barrier-to-autointegration factor B-like and serine/arginine-rich splicing factor 1 A-like. When the cholesterol of the lipid rafts from the membrane was depleted by methyl-β-cyclodextrin (MβCD), BmNPV entry into BmN cells was blocked. However, supplying cholesterol into the medium rescued the BmNPV infection ability. These results show that membrane lipid rafts may be the active regions for the entry of BmNPV into cells, and the components of membrane lipid rafts may be candidate targets for improving the resistance of the silkworm to BmNPV.

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    Authors and Affiliations

    1. School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China

      Xiaolong Hu, Min Zhu, Zi Liang, Dhiraj Kumar, Fei Chen, Liyuan Zhu, Sulan Kuang, Renyu Xue, Guangli Cao & Chengliang Gong

    2. National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China

      Xiaolong Hu, Renyu Xue, Guangli Cao & Chengliang Gong

    3. Institute of Agricultural Biotechnology and Ecological Research, Soochow University, Suzhou, 215123, China

      Xiaolong Hu, Renyu Xue, Guangli Cao & Chengliang Gong

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    1. Xiaolong Hu
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    2. Min Zhu
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    3. Zi Liang
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    Correspondence to Chengliang Gong.

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    This article does not contain any studies with human participants or animals performed by any of the authors.

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    This study was funded by the Natural Science Foundation of Jiangsu Higher Education Institutions of China (15KJB230002), the Natural Science Foundation of Jiangsu Province (BK20140324), the National Natural Science Foundation of China (31072085) and a project funded by the Priority Academic Program of Development of Jiangsu Higher Education Institutions.

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    Communicated by Q. Xia.

    X. Hu and M. Zhu contributed equally to this work.

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    Hu, X., Zhu, M., Liang, Z. et al. Proteomic analysis of BmN cell lipid rafts reveals roles in Bombyx mori nucleopolyhedrovirus infection. Mol Genet Genomics 292, 465–474 (2017). https://doi.org/10.1007/s00438-016-1284-y

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