Genomic and Proteomic Analysis of Thirty-Nine Structural Proteins of Shrimp White Spot Syndrome Virus

Abstract

White spot syndrome virus (WSSV) virions were purified from the hemolymph of experimentally infected crayfish Procambarus clarkii, and their proteins were separated by 8 to 18% gradient sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) to give a protein profile. The visible bands were then excised from the gel, and following trypsin digestion of the reduced and alkylated WSSV proteins in the bands, the peptide sequence of each fragment was determined by liquid chromatography-nano-electrospray ionization tandem mass spectrometry (LC-nanoESI-MS/MS) using a quadrupole/time-of-flight mass spectrometer. Comparison of the resulting peptide sequence data against the nonredundant database at the National Center for Biotechnology Information identified 33 WSSV structural genes, 20 of which are reported here for the first time. Since there were six other known WSSV structural proteins that could not be identified from the SDS-PAGE bands, there must therefore be a total of at least 39 (33 ؉ 6) WSSV structural protein genes. Only 61.5% of the WSSV structural genes have a polyadenylation signal, and preliminary analysis by 3 rapid amplification of cDNA ends suggested that some structural protein genes produced mRNA without a poly(A) tail. Microarray analysis showed that gene expression started at 2, 6, 8, 12, 18, 24, and 36 hpi for 7, 1, 4, 12, 9, 5, and 1 of the genes, respectively. Based on similarities in their time course expression patterns, a clustering algorithm was used to group the WSSV structural genes into four clusters. Genes that putatively had common or similar roles in the viral infection cycle tended to appear in the same cluster.

Differentially expressed genes in Penaeus monodon hemocytes following infection with yellow head virus

“…In contrast, the responses of shrimp to YHV infection at the molecular level are currently poorly described. WSSV is an enveloped, ellipsoid and large double-stranded DNA virus (5), while YHV is an enveloped, rod-shaped and single-stranded RNA virus (6). As YHV contains a single-stranded RNA genome, the molecular basis for shrimp response to YHV might be somewhat different than that to the DNA virus WSSV.…”

Fish reovirus GCReV-109 VP33 protein elicits protective immunity in rare minnows

“…The Western blot result was in accord with SDS-PAGE analysis, which confirmed that band 5 of the SDS-PAGE gel was derived from GCReV-109 virions and corresponded to the VP33 protein. We therefore conclude that VP33 is a major structural protein of GCReV-109 virions [27][28][29].…”

Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody

“…The ORF WSSV388 encodes a putative 786 amino acid protein (88 kDa). A recent proteomic analysis (Tsai et al, 2004) confirms the existence of this protein which was named VP75 based on its migration in an SDS-PAGE. PROSITE database analysis indicated that WSSV388 contained a putative immunoglobulin/major histocompatibility complex (IG MHC) protein signature motif.…”

“…Only a few major structural proteins (Van Hulten et al, 2000a,b;Chen et al, 2002;Zhang et al, 2002) and enzymes (Tsai et al, 2000;Li et al, 2004) date. SDS-PAGE and 2-D electrophoresis were combined with mass spectrometry, which led to the successful identification of new structural proteins that have been used to study the WSSV proteome Tsai et al, 2004;Zhang et al, 2004). However, the role of most of these viral proteins and the molecular mechanism of infection are still not known.…”