Characterization and interactome study of white spot syndrome virus envelope protein VP11.
White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570), and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.
Department of Earth and Life Science, University of Taipei, Taipei, Taiwan.
Wang-Jing
WJ
Liu
Department of Molecular Biotechnology, Da-Yeh University, Changhua, Taiwan.
Hui-Jui
HJ
Shiung
Institute of Bioinformatics and Biosignal Transduction, National Cheng Kung University, Tainan, Taiwan.
Chu-Fang
CF
Lo
Institute of Marine Biology, National Taiwan Ocean University, Keelung, Taiwan ; Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan.
Jiann-Horng
JH
Leu
Department of Food Science, National Quemoy University, Kinmen, Taiwan.
Ying-Jang
YJ
Lai
Department of Molecular Biotechnology, Da-Yeh University, Changhua, Taiwan.
Tai-Lin
TL
Lee
Department of Molecular Biotechnology, Da-Yeh University, Changhua, Taiwan.
Wei-Tung
WT
Huang
Institute of Zoology, National Taiwan University, Taipei, Taiwan.
Guang-Hsiung
GH
Kou
Department of Molecular Biotechnology, Da-Yeh University, Changhua, Taiwan.
Yun-Shiang
YS
Chang
D015967:Gene Expression Regulation, Viral; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D011485:Protein Binding; D055503:Protein Multimerization; D015203:Reproducibility of Results; D013997:Time Factors; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014771:Virion / Q000378:metabolism; D046848:White spot syndrome virus 1 / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D014759:Viral Envelope Proteins