Characterization of White Spot Syndrome Virus Envelope Protein VP51A and Its Interaction with Viral Tegument Protein VP26
Characterization of White Spot Syndrome Virus Envelope Protein VP51A and Its Interaction with Viral Tegument Protein VP26
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DOI：
10.1128/jvi.01238-08
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发表时间：
2008-12-15
影响因子：
5.4
通讯作者：
Lo, Chu-Fang
中科院分区：
文献类型：
--
作者：
Chang, Yun-Shiang;Liu, Wang-Jing;Lo, Chu-Fang
在这项研究中，我们鉴定了一种新的白斑综合征病毒(WSSV)结构蛋白VP51A(WSSV-TW开放阅读框架294)，它是从先前的质谱学研究中鉴定出来的。时间转录分析表明，vp51a在WSSV感染的晚期有表达。基因结构分析表明，vp51a的转录起始点位于翻译起始密码子上游135个碱基。Poly(A)添加信号与翻译终止密码子TAA重叠，Poly(A)尾部位于TAA下游23bp。蛋白质组分的Western印迹分析和免疫电子显微镜均表明VP51a是一种病毒包膜蛋白。从WSSV病毒粒子中提取的总蛋白的Western blotting检测到一条与预测的51 kDa质量接近的条带，但最强的信号在72 kDa左右。我们得出结论，这条72 kDa的条带实际上是全长的VP51a蛋白。膜拓扑学分析表明，VP51A72-kDa蛋白是一种II型跨膜蛋白，其N端有一个高度疏水的跨膜区，C端暴露在病毒粒子表面。免疫共沉淀、共定位和酵母双杂交实验表明，VP51a与VP26直接结合，与VP28间接结合，VP26作为连接蛋白参与VP51a-VP26VP28复合体的形成。
In this study, we characterize a novel white spot syndrome virus (WSSV) structural protein, VP51A (WSSV-TW open reading frame 294), identified from a previous mass spectrometry study. Temporal-transcription analysis showed that vp51A is expressed in the late stage of WSSV infection. Gene structure analysis showed that the transcription initiation site of vp51A was 135 bp upstream of the translation start codon. The poly(A) addition signal overlapped with the translation stop codon, TAA, and the poly(A) tail was 23 bp downstream of the TAA. Western blot analysis of viral protein fractions and immunoelectron microscopy both suggested that VP51A is a viral envelope protein. Western blotting of the total proteins extracted from WSSV virions detected a band that was close to the predicted 51-kDa mass, but the strongest signal was around 72 kDa. We concluded that this 72-kDa band was in fact the full-length VP51A protein. Membrane topology assays demonstrated that the VP51A 72-kDa protein is a type II transmembrane protein with a highly hydrophobic transmembrane domain on its N terminus and a C terminus that is exposed on the surface of the virion. Coimmunoprecipitation, colocalization, and yeast two-hybrid assays revealed that VP51A associated directly with VP26 and indirectly with VP28, with VP26 acting as a linker protein in the formation of a VP51A-VP26VP28 complex.