Studies on the Interactions of White Spot Syndrome Virus Structural Proteins and Penaeus monodon Cellular Proteins, PmRACK1, PmRab5B, and PmRab6A

• Main Authors: Te-Chang Liao, 廖德昌
• Other Authors: Yun-Shiang Chang
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/51206809194905610565

碩士 === 大葉大學 === 分子生物科技學系碩士班 === 102 === White spot syndrome virus (WSSV, genus Whispovirus, family Nimaviridae) is a wide occurring virus which attacks cultured shrimp and many other crustaceans and has caused huge economic losses for the global shrimp farming industry for more than a decade. Until now effective methods for dealing with the disease caused by WSSV infection is not available. Recently several host cellular proteins related to WSSV infection such as Rab6, Rab7, gC1qR (receptor for the globular heads of C1q), F1ATP synthase β subunit, RACK1 (receptor for activated protein kinase C1) and CBP (chitin binding protein) had been reported. In this study, full length of PmRab5B (Penaeus monodon Rab5B), PmRab6A (P. monodon Rab6A) genes were firstly cloned. The pmRab5B, pmRab6A, and one other known WSSV infection related P. monodon cellular protein PmRACK1 (P. monodon receptor for activated protein kinase C1) were used as bait for screening of their interaction WSSV structural proteins on a previously prepared yeast two-hybrid library. Yeast two-hybrid screening revealed that PmRab5B, PmRab6A and PmRACK1 interacted with 31, 36, and 21 viral structure proteins, respectively. The following co-immunoprecipitation assays confirmed the interactions of PmRab5B with WSSV VP31; PmRab6A with WSSV VP53A, VP41A, VP51A and VP36A; PmRACK1 with WSSV VP11, VP39 and VP36A. The anti-WSSV assays performed on Litopenaeus vannamei by a PmRACK1 interaction WSSV structural protein, the VP11, showed an obvious effect. The recombinant VP11 protein treated shrimp were with greater than 30% survival rate than the untreated ones. Results of this study would contribute in understanding of the mechanisms of WSSV infection and also give valuable hints in developing of anti-WSSV strategies. Key Words: WSSV, Penaeus monodon, virus-host interaction, protein-protein interaction, VP11