Research on the Entry of White Spot Syndrome Virus (WSSV) into Host Cells---Analysis the Envelope Protein Complex

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Project title

Code/計畫編號

NSC99-2311-B019-001

Translated Name/計畫中文名

白點症病毒入侵寄主細胞機轉之研究---封套蛋白複合體之解析

Project Coordinator/計畫主持人

Li-Li Chen

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=2131723

Year

2010

Start date/計畫起

01-08-2010

Expected Completion/計畫迄

31-07-2011

Bugetid/研究經費

900千元

ResearchField/研究領域

生物科學

白點症病毒是造成台灣以及世界其他地區養殖蝦類嚴重死亡的病原體，儘管目前在白點症病毒的基因體和蛋白質體學方面的研究相當多，然而對於病毒如何入侵寄主細胞的機制卻仍不清楚。在過去的研究中，我們利用酵母菌雙雜合法篩選能與白點症病毒封套蛋白VP53A 結合的蛋白質，發現草蝦幾丁質結合蛋白能與之結合，並驗證確定草蝦幾丁質結合蛋白位在細胞膜上，分子量約為34 kDa。我們利用重組VP53A 和幾丁質結合蛋白對白蝦進行活體感染阻斷試驗，發現白蝦的死亡率可抑制20～40％。再進一步以專一於幾丁質結合蛋白基因的dsRNA 進行基因減弱，發現幾丁質結合基因減弱處理組的死亡率為0％，顯示幾丁質結合蛋白與病毒入侵寄主細胞有關。我們最近利用酵母菌雙雜合法篩選能與草蝦幾丁質結合蛋白結合的病毒蛋白，發現至少有11 個病毒封套蛋白能與其結合，也同時發現一新穎蛋白質—葡萄醣運輸蛋白—可與VP53A 結合，因此我們推測VP53A 等11 個病毒封套蛋白可能形成一複合體，參與了病毒入侵機制。因此在本期的計畫中，我們將由解開病毒封套蛋白複合體的結構與作用模式著手，冀望能解開白點症病毒入侵寄主細胞機轉，並選出最好的標的物，開發出最有效的抗病毒策略應用於產業界。"White spot syndrome virus (WSSV) can cause the most serious viral disease of shrimp and has a wide host range among crustaceans. Although researches showed a lot about its genome and structure, information concerning the mechanism of how WSSV infects cells was lacking. In the previous study, WSSV envelope protein, VP53A, was identified to interact with Penaeus monodon chitin-binding protein (PmCBP). PmCBP, renamed as CBP in the later study, was confirmed to locate on the cell surface of the host cell, and its molecular weight was about 34 kDa. In the in vivo infection blocking assay, both rVP53A and rCBP that were produced by Esherichia coli can promote resp. a 40% and 20% survival rate of the shrimp which were challenged by WSSV. Recent study using specific dsRNA to knock-down CBP gene indicated that cumulative mortality of CBP-dsRNA treated group was reduced to 0%. Those findings suggested that CBP was involved in WSSV infection. Furthermore, a yeast-two-hybrid result revealed that CBP could interact with at least 11 WSSV envelope proteins. Moreover, a new gene, glucose transporter (Gluct1), was also found to interact with VP53A. Thus, we speculate that these 11 WSSV envelope proteins may form a complex to mediate virus entry. Therefore, a project based on revealing the viral envelope complex was proposed. We hope the study can help to understand the mechanism of how WSSV enter host cells, and the result can ultimately help to develop the most efficient anti-viral strategy against WSSV."

Keyword(s)

白點症病毒
封套蛋白
草蝦幾丁質結合蛋白
葡萄醣運輸蛋白
複合體
抗病毒策略
WSSV
Gluct1
envelope protein
PmCBP or CBP
complex
anti-viral strategy

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Research on the Entry of White Spot Syndrome Virus (WSSV) into Host Cells---Analysis the Envelope Protein Complex

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