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Published online ahead of print on 4 March 2009 as doi:10.1099/vir.0.008912-0
Journal of General Virology 2009;90:1483.

A more recent version of this article appeared on June 1, 2009 J Gen Virol (2009), DOI 10.1099/vir.0.008912-0
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Viral ubiquitin ligase WSSV222 is required for efficient white spot syndrome virus (WSSV) replication in shrimp

Fang He1, Syed Musthaq Syed1, A. S. Sahul Hameed2 and Jimmy Kwang3,4

1 Temasek Life Sciences Laboratory;
2 OIE Reference Laboratory for WTD, Department of Zoology, C. Abdul Hakeem College;
3 Animal Health Biotechnology, Temasek Life Sciences Laboratory

4 E-mail: kwang{at}tll.org.sg

The E3 ligase WSSV222 of white spot syndrome virus (WSSV) is involved in the anti-apoptosis regulation by ubiquitin-mediated degradation on TSL (tumor suppressor like protein), a shrimp tumor suppressor. In the present study, WSSV222 expression was silenced by specific siRNA in Sf9 and BHK cells. Based on the results of the in vitro silencing, WSSV-challenged shrimp were treated with anti-222 siRNA to knockdown WSSV222. The survival rate and the efficiency of WSSV replication were assessed to evaluate the efficacy of anti-222 siRNA to regulate WSSV infection in shrimp. The anti-222 siRNA reduced the cumulative mortality in shrimp challenged with 103 copies of WSSV and delayed the mean time to death in shrimp challenged with the higher dosage of 106 copies. The results of real time quantitative PCR showed that virus replication was delayed and reduced in the WSSV-challenged shrimp treated with anti-222 siRNA in comparison to the challenged shrimp treated with random siRNA. Co-immunoprecipitation assays revealed that WSSV222 silencing inhibited the degradation of TSL in WSSV-challenged shrimp and the results indicate the requirement of WSSV222 for efficient replication of WSSV in shrimp.

Received 18 November 2008; accepted 23 January 2009.




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