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Journal of Virology, March 2006, p. 3021-3029, Vol. 80, No. 6
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.6.3021-3029.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of the Nucleocapsid, Tegument, and Envelope Proteins of the Shrimp White Spot Syndrome Virus Virion

Jyh-Ming Tsai,1 Han-Ching Wang,1 Jiann-Horng Leu,1 Andrew H.-J. Wang,2 Ying Zhuang,3 Peter J. Walker,4 Guang-Hsiung Kou,1* and Chu-Fang Lo1*

Institute of Zoology, National Taiwan University, Taipei, Taiwan,1 Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan,2 Department of Biological Sciences, National University of Singapore, Singapore,3 CSIRO Livestock Industries, Queensland Bioscience Precinct, St. Lucia, Australia4

Received 15 September 2005/ Accepted 5 December 2005

The protein components of the white spot syndrome virus (WSSV) virion have been well established by proteomic methods, and at least 39 structural proteins are currently known. However, several details of the virus structure and assembly remain controversial, including the role of one of the major structural proteins, VP26. In this study, Triton X-100 was used in combination with various concentrations of NaCl to separate intact WSSV virions into distinct fractions such that each fraction contained envelope and tegument proteins, tegument and nucleocapsid proteins, or nucleocapsid proteins only. From the protein profiles and Western blotting results, VP26, VP36A, VP39A, and VP95 were all identified as tegument proteins distinct from the envelope proteins (VP19, VP28, VP31, VP36B, VP38A, VP51B, VP53A) and nucleocapsid proteins (VP664, VP51C, VP60B, VP15). We also found that VP15 dissociated from the nucleocapsid at high salt concentrations, even though DNA was still present. These results were confirmed by CsCl isopycnic centrifugation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-nanoelectrospray ionization-tandem mass spectrometry, by a trypsin sensitivity assay, and by an immunogold assay. Finally, we propose an assembly process for the WSSV virion.

* Corresponding author. Mailing address: Institute of Zoology, National Taiwan University, Taipei 106, Taiwan, Republic of China. Phone: 886-2-23633562. Fax: 886-2-23638179. E-mail for Guang-Hsiung Kou: ghkou{at}ntu.edu.tw. E-mail for Chu-Fang Lo: gracelow{at}ntu.edu.tw.

Journal of Virology, March 2006, p. 3021-3029, Vol. 80, No. 6
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.6.3021-3029.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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