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Journal of Virology, November 2006, p. 10734-10742, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.00349-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

PmRab7 Is a VP28-Binding Protein Involved in White Spot Syndrome Virus Infection in Shrimp

Kallaya Sritunyalucksana,1,2 Wanphen Wannapapho,1,2 Chu Fang Lo,3 and Timothy W. Flegel2*

National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani 12120, Thailand,1 Centex Shrimp, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand,2 Institute of Zoology, National Taiwan University, Taipei, Taiwan, Republic of China3

Received 23 February 2006/ Accepted 31 July 2006

Our aim was to isolate and characterize white spot syndrome virus (WSSV)-binding proteins from shrimp. After a blot of shrimp hemocyte membrane proteins was overlaid with a recombinant WSSV envelope protein (rVP28), the reactive bands on the blot were detected using anti-VP28 antibody. Among three membrane-associated molecules identified by liquid chromatography-tandem mass spectrometry, there was a 25-kDa protein that bound to both rVP28 and WSSV. Since it had a primary structure with high homology to the small GTP-binding protein Rab7, we named it Penaeus monodon Rab7 (PmRab7). The full-length PmRab7 cDNA was obtained, and results from a glutathione S-transferase pull-down assay confirmed specific binding to rVP28. Reverse transcriptase PCR analysis revealed PmRab7 expression in many tissues, and real-time PCR analysis revealed that expression was constitutive. Binding of PmRab7 to rVP28 or WSSV occurred in a dose-dependent manner and was inhibited by anti-Rab7 antibody. In an in vivo neutralization assay, the number of dead shrimp after challenge with WSSV plus PmRab7 (15%) or WSSV plus anti-Rab7 antibody (5%) was significantly lower than after challenge with WSSV alone (95%). In contrast to the WSSV-injected group, shrimp injected with WSSV plus PmRab7 or WSSV plus anti-Rab7 showed no WSSV-type histopathology. We conclude that PmRab7 is involved in WSSV infection in shrimp. This is the first study to identify a shrimp protein that binds directly to a major viral envelope protein of WSSV.

* Corresponding author. Mailing address: Center of Excellence in Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Phyathai, Bangkok 10400, Thailand. Phone: 66-02-201-5878. Fax: 66-02-354-7344. E-mail: sctwf{at}mahidol.ac.th.

Journal of Virology, November 2006, p. 10734-10742, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.00349-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • He, F., Syed, S. M., Hameed, A. S. S., Kwang, J. (2009). Viral ubiquitin ligase WSSV222 is required for efficient white spot syndrome virus replication in shrimp. J. Gen. Virol. 90: 1483-1490 [Abstract] [Full Text]  
  • Wang, H.-C., Wang, H.-C., Ko, T.-P., Lee, Y.-M., Leu, J.-H., Ho, C.-H., Huang, W.-P., Lo, C.-F., Wang, A. H.-J. (2008). White spot syndrome virus protein ICP11: A histone-binding DNA mimic that disrupts nucleosome assembly. Proc. Natl. Acad. Sci. USA 105: 20758-20763 [Abstract] [Full Text]  
  • Tang, X., Wu, J., Sivaraman, J., Hew, C. L. (2007). Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship. J. Virol. 81: 6709-6717 [Abstract] [Full Text]  


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