Proteomic analysis of BmN cell lipid rafts reveals roles in Bombyx mori nucleopolyhedrovirus infection.
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.
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China.|Institute of Agricultural Biotechnology and Ecological Research, Soochow University, Suzhou, 215123, China.
Xiaolong
X
Hu
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Min
M
Zhu
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Zi
Z
Liang
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Dhiraj
D
Kumar
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Fei
F
Chen
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Liyuan
L
Zhu
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.
Sulan
S
Kuang
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China.|Institute of Agricultural Biotechnology and Ecological Research, Soochow University, Suzhou, 215123, China.
Renyu
R
Xue
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China.|Institute of Agricultural Biotechnology and Ecological Research, Soochow University, Suzhou, 215123, China.
Guangli
G
Cao
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China. gongcl@suda.edu.cn.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China. gongcl@suda.edu.cn.|Institute of Agricultural Biotechnology and Ecological Research, Soochow University, Suzhou, 215123, China. gongcl@suda.edu.cn.
Chengliang
C
Gong
D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000821:virology*; D002460:Cell Line; D002784:Cholesterol / Q000737:chemistry; D019295:Computational Biology; D020869:Gene Expression Profiling; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry; D008055:Lipids / Q000737:chemistry; D013058:Mass Spectrometry; D021962:Membrane Microdomains / Q000737:chemistry*; D058977:Molecular Sequence Annotation; D017924:Nucleopolyhedroviruses / Q000502:physiology*; D040901:Proteomics
D018841:HSP90 Heat-Shock Proteins; D019476:Insect Proteins; D008055:Lipids; D002784:Cholesterol
Bombyx mori; Bombyx mori nucleopolyhedrovirus; Cholesterol; Lipid rafts