Purification and characterization of White Spot Syndrome Virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii

Huang, Canhua; Li-Ren, Zhang; Zhang, Jianhong; Xiao, Lanbo; Wu, Qingping; Chen, Di-Hua; Li, Joseph K.-K.

doi:10.1016/s0168-1702(01)00247-7

Cited by 88 publications

(66 citation statements)

References 11 publications

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“…Moreover, some MALDI-TOF MS results were confirmed. Although the sequence coverage of bands 11,13,14,16,19, and 23 to the predicted proteins of the vp281, vp292, p22, vp384, vp544, and vp800 genes were lower, at least 10 successive amino acid sequences were matched. These sequence tags should be specific.…”

Section: Discussionmentioning

confidence: 96%

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Proteomic Analysis of Shrimp White Spot Syndrome Viral Proteins and Characterization of a Novel Envelope Protein VP466

Huang

Zhang

Lin

et al. 2002

Molecular & Cellular Proteomics

127

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White spot syndrome virus (WSSV) is at present one of the major pathogens in shrimp culture worldwide. The complete genome of this virus has been sequenced recently. To identify the structural and functional proteins of WSSV, the purified virions were separated by SDS-PAGE. Twenty-four protein bands were excised, in-gel digested with trypsin, and subjected to matrix-assisted laser desorption ionization-time of flight mass spectrometry and electrospray ionization tandem mass spectrometry, respectively. White spot syndrome virus (WSSV), 1 considered to be a new virus (1), is one of the major pathogens in cultured penaeid shrimp. First appearing in the 1990s in Taiwan, WSSV has spread rapidly to shrimp-farming areas around the world causing large economic losses. The virus has a broad host range, including other invertebrate aquatic organisms, such as crab and crayfish (2, 3). The WSSV particles are nonoccluded and bacilliform in shape with double envelopes (4). In 1997, the WSSV genomic DNA was purified successfully from Penaeus japonicus shrimp in China (5), and the genomic DNA and cDNA libraries were constructed. WSSV genome contains a 305-kb double-stranded circular DNA (6), which is longer than the 293-kb genome isolated from Penaeus monodon shrimps in Thailand (7). Approximately 180 open reading frames (ORFs) of 50 amino acids or more were revealed by the analysis of the WSSV genomic DNA sequences (6). However, in contrast to the insect baculovirus, one of the best studied viruses, only a few genes from WSSV have been reported (8).With the completion of the WSSV genomic DNA sequence, research has now focused on the functional analysis of the gene products. Essential to the functional analysis is to identify the proteins expressed in WSSV. To this end, a proteomic approach using mass spectrometry has been proven to be the most effective technology for the identification of proteins (9). Recently, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) utilizing a quadrupole time-of-flight (Q-TOF) mass spectrometer have been used as tools for the characterization of proteins because of their high sensitivity and throughput (10).In this communication, the WSSV proteins separated by SDS-PAGE were analyzed using MALDI-TOF MS and ESI-MS/MS (Q-TOF), respectively. The resulting mass spectrometric data were searched against the theoretical ORF database of WSSV. One of the newly retrieved genes, vp466 gene, was further characterized. EXPERIMENTAL PROCEDURES Proliferation and Purification of Shrimp WSSVProliferation and Purification of WSSV Virion-The infected tissues from penaeid shrimp P. monodon (e.g. gill, stomach, midgut, etc.) From the ‡Department

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Section: Discussionmentioning

confidence: 96%

“…The resulting pellets were resuspended in TNE. Virus samples were examined under a transmission electron microscope (JEOL 100 CXII) for purity (11).…”

Section: Proliferation and Purification Of Shrimp Wssvmentioning

confidence: 99%

Proteomic Analysis of Shrimp White Spot Syndrome Viral Proteins and Characterization of a Novel Envelope Protein VP466

Huang

Zhang

Lin

et al. 2002

Molecular & Cellular Proteomics

127

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“…It infects several species of penaeid shrimp cultivated in the Eastern/Western Hemispheres (Lu et al 1997, Corsin et al 2001, in addition to a wide range of other decapods and crustaceans, including crabs (Chen et al 2000, Sahul Hameed et al 2001) and fresh water crayfish (Huang et al 2001, Jiravanichpaisal et al 2001. Several studies have confirmed that many suspected decapod carriers of WSSV can transmit the virus to Penaeus monodon (Wongteerasupaya et al 1996, Chang et al 1998, Wang et al 1998.…”

Section: Introductionmentioning

confidence: 98%

White spot syndrome virus (WSSV) infectivity for Artemia at different developmental stages

Zhang²,

Chen³

et al. 2003

Dis. Aquat. Org.

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White spot syndrome virus (WSSV) is a major pathogen of cultivated shrimp, but its host range includes a large number of crustaceans. In this investigation, Artemia franciscana was tested for susceptibility to WSSV by the oral route. Both instars and adults were challenged, and the presence of WSSV was followed through to reproductive cysts and offspring using PCR. WSSV caused a much lower cumulative mortality in Artemia than in cultivated shrimp by 10 d post-challenge. Instars, adults and reproductive cysts were PCR positive. However, the virus was undetectable by PCR in nauplii that had hatched from PCR-positive reproductive cysts. The data indicate that WSSV or WSSV genomic DNA can be vertically transmitted from WSSV-PCR-positive instars to reproductive cysts, but this DNA is removed during hatching.

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“…Among them, VP28, VP26, VP24, and VP19 are the four major viral envelope proteins (20,21); these proteins form a multiprotein complex together with some low-abundance proteins (22)(23)(24)(25)(26). In addition to the importance of this envelope protein complex in virus assembly, it is thought to be an "infectome" that functions in cell recognition during infection (24,27).…”

Section: Importancementioning

confidence: 99%

VP24 Is a Chitin-Binding Protein Involved in White Spot Syndrome Virus Infection

Han

et al. 2016

J Virol

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Oral ingestion is the major route of infection for the white spot syndrome virus (WSSV). However, the mechanism by which virus particles in the digestive tract invade host cells is unknown. In the present study, we demonstrate that WSSV virions can bind to chitin through one of the major envelope proteins (VP24). Mutagenesis analysis indicated that amino acids (aa) 186 to 200 in the C terminus of VP24 were required for chitin binding. Moreover, the P-VP24 186 -200 peptide derived from the VP24 chitin binding region significantly inhibited the VP24-chitin interaction and the WSSV-chitin interaction, implying that VP24 participates in WSSV binding to chitin. Oral inoculation experiments showed that P-VP24 186 -200 treatment reduced the number of virus particles remaining in the digestive tract during the early stage of infection and greatly hindered WSSV proliferation in shrimp. These data indicate that binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection and provide new ideas for preventing WSSV infection in shrimp farms. IMPORTANCEIn this study, we show that WSSV can bind to chitin through the envelope protein VP24. The chitin-binding domain of VP24 maps to amino acids 186 to 200 in the C terminus. Binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection. These findings not only extend our knowledge of WSSV infection but also provide new insights into strategies to prevent WSSV infection in shrimp farms. W hite spot syndrome virus (WSSV) is an enveloped doublestranded DNA (dsDNA) virus belonging to the genus Whispovirus, family Nimaviridae (1). WSSV has caused enormous economic losses to the shrimp farming industry since 1993. Infectious WSSV virions are typically 250 to 380 nm in length and 120 to 150 nm in diameter (2, 3), and they contain a genome of ϳ300 kb that encodes ϳ180 proteins (4, 5).Ingestion of WSSV-infected sick or dead shrimp has been accepted as the major route of natural infection due to the cannibalistic nature of shrimp (6-11). Therefore the digestive tract of shrimp may be a primary site of infection. The digestive tract of shrimp is composed of the esophagus, stomach, midgut, and hindgut. The esophagus, stomach, and hindgut possess a chitinous lining, which is not present in the midgut (12, 13). Instead, the midgut epithelium is generally lined with the peritrophic membrane (PM), which is a noncellular structure surrounding the food bolus. The PM is composed of regularly arranged chitin fibrils embedded in a matrix of proteins, proteoglycans, and mucopolysaccharides (14-17). Therefore, WSSV must cross the PM in the midgut or the chitinous lining in the other parts of the digestive tract to traverse the basal membranes and reach the host cells. We speculate that the interaction between WSSV and chitin may be important for WSSV infection in the shrimp digestive tract.More than 40 WSSV structural proteins have been identified using proteomic methods to date (18)(19)(20). Among them, VP28,...

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Purification and characterization of White Spot Syndrome Virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii

Cited by 88 publications

References 11 publications

Proteomic Analysis of Shrimp White Spot Syndrome Viral Proteins and Characterization of a Novel Envelope Protein VP466

Proteomic Analysis of Shrimp White Spot Syndrome Viral Proteins and Characterization of a Novel Envelope Protein VP466

White spot syndrome virus (WSSV) infectivity for Artemia at different developmental stages

VP24 Is a Chitin-Binding Protein Involved in White Spot Syndrome Virus Infection

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