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

2004 ◽  
Vol 78 (20) ◽  
pp. 11360-11370 ◽  
Author(s):  
Jyh-Ming Tsai ◽  
Han-Ching Wang ◽  
Jiann-Horng Leu ◽  
He-Hsuan Hsiao ◽  
Andrew H.-J. Wang ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
Expression Patterns ◽  
Sodium Dodecyl ◽  
Peptide Sequence ◽  
White Spot ◽  
Structural Proteins ◽  
Structural Protein ◽  
Structural Genes ◽  
Sds Page ◽  
White Spot Syndrome

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 sulfate-polyacrylamide 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.

scholarly journals Identification of the Nucleocapsid, Tegument, and Envelope Proteins of the Shrimp White Spot Syndrome Virus Virion

2006 ◽  
Vol 80 (6) ◽  
pp. 3021-3029 ◽  
Author(s):  
Jyh-Ming Tsai ◽  
Han-Ching Wang ◽  
Jiann-Horng Leu ◽  
Andrew H.-J. Wang ◽  
Ying Zhuang ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Envelope Proteins ◽  
White Spot ◽  
Structural Proteins ◽  
Nucleocapsid Proteins ◽  
Tegument Proteins ◽  
White Spot Syndrome ◽  
Protein Components

ABSTRACT The protein components of the white spot syndrome virus (WSSV) virion have been well established by proteomic methods, and at least 39 structural proteins are currently known. However, several details of the virus structure and assembly remain controversial, including the role of one of the major structural proteins, VP26. In this study, Triton X-100 was used in combination with various concentrations of NaCl to separate intact WSSV virions into distinct fractions such that each fraction contained envelope and tegument proteins, tegument and nucleocapsid proteins, or nucleocapsid proteins only. From the protein profiles and Western blotting results, VP26, VP36A, VP39A, and VP95 were all identified as tegument proteins distinct from the envelope proteins (VP19, VP28, VP31, VP36B, VP38A, VP51B, VP53A) and nucleocapsid proteins (VP664, VP51C, VP60B, VP15). We also found that VP15 dissociated from the nucleocapsid at high salt concentrations, even though DNA was still present. These results were confirmed by CsCl isopycnic centrifugation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-nanoelectrospray ionization-tandem mass spectrometry, by a trypsin sensitivity assay, and by an immunogold assay. Finally, we propose an assembly process for the WSSV virion.

scholarly journals The Unique Stacked Rings in the Nucleocapsid of the White Spot Syndrome Virus Virion Are Formed by the Major Structural Protein VP664, the Largest Viral Structural Protein Ever Found

2005 ◽  
Vol 79 (1) ◽  
pp. 140-149 ◽  
Author(s):  
Jiann-Horng Leu ◽  
Jyh-Ming Tsai ◽  
Han-Ching Wang ◽  
Andrew H.-J. Wang ◽  
Chung-Hsiung Wang ◽  
...  
Keyword(s):  
Immunoelectron Microscopy ◽  
Time Course ◽  
White Spot Syndrome Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
White Spot ◽  
Sequencing Analysis ◽  
Structural Protein ◽  
Reading Frame ◽  
White Spot Syndrome

ABSTRACT One unique feature of the shrimp white spot syndrome virus (WSSV) genome is the presence of a giant open reading frame (ORF) of 18,234 nucleotides that encodes a long polypeptide of 6,077 amino acids with a hitherto unknown function. In the present study, by applying proteomic methodology to analyze the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of purified WSSV virions by liquid chromatography-mass spectrometry (LC-MS/MS), we found that this giant polypeptide, designated VP664, is one of the viral structural proteins. The existence of the corresponding 18-kb transcript was confirmed by sequencing analysis of reverse transcription-PCR products, which also showed that vp664 was intron-less. A time course analysis showed that this transcript was actively transcribed at the late stage, suggesting that this gene product should contribute primarily to the assembly and morphogenesis of the virion. Several polyclonal antisera against this giant protein were prepared, and one of them was successfully used for immunoelectron microscopy analysis to localize the protein in the virion. Immunoelectron microscopy with a gold-labeled secondary antibody showed that the gold particles were regularly distributed around the periphery of the nucleocapsid with a periodicity that matched the characteristic stacked ring subunits that appear as striations. From this and other evidence, we argue that this giant ORF in fact encodes the major WSSV nucleocapsid protein.

scholarly journals Proteomic Analysis of the Major Envelope and Nucleocapsid Proteins of White Spot Syndrome Virus

2006 ◽  
Vol 80 (21) ◽  
pp. 10615-10623 ◽  
Author(s):  
Xixian Xie ◽  
Limei Xu ◽  
Feng Yang
Keyword(s):  
Posttranslational Modifications ◽  
White Spot Syndrome Virus ◽  
Procambarus Clarkii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
White Spot ◽  
Structural Proteins ◽  
Nucleocapsid Proteins ◽  
Flight Mass Spectrometry ◽  
White Spot Syndrome

ABSTRACT White spot syndrome virus (WSSV) virions were purified from the tissues of infected Procambarus clarkii (crayfish) isolates. Pure WSSV preparations were subjected to Triton X-100 treatment to separate into the envelope and nucleocapsid fractions, which were subsequently separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major envelope and nucleocapsid proteins were identified by either matrix-assisted laser desorption ionization-time of flight mass spectrometry or defined antibody. A total of 30 structural proteins of WSSV were identified in this study; 22 of these were detected in the envelope fraction, 7 in the nucleocapsid fraction, and 1 in both the envelope and the nucleocapsid fractions. With the aid of specific antibodies, the localizations of eight proteins were further studied. The analysis of posttranslational modifications revealed that none of the WSSV structural proteins was glycosylated and that VP28 and VP19 were threonine phosphorylated. In addition, far-Western and coimmunoprecipitation experiments showed that VP28 interacted with both VP26 and VP24. In summary, the data obtained in this study should provide an important reference for future molecular studies of WSSV morphogenesis.

scholarly journals In-Vivo Comparative Virulence of Different White Spot Syndrome Virus Isolates In Penaeus Vannamei And Whole Genome Comparison Analysis

2021 ◽  
Author(s):  
Álvaro Hernán Hernández-Montiel ◽  
Manfred Weidmann ◽  
Michael Bekaert ◽  
Kristina Ulrich ◽  
Jessica Benkaroun ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
White Spot ◽  
White Shrimp ◽  
Genome Comparison ◽  
Structural Protein ◽  
High Virulence ◽  
Multiple Routes ◽  
White Spot Syndrome ◽  
Virus Isolates

Abstract White Spot Syndrome Virus (WSSV) infects several economically important aquacultural species, causing significant losses to the industry. This virus belongs to the Nimaviridae family, and has a dsDNA genome ranging from 257 to 309 kb (more than 20 isolate genomes fully sequenced and published to date). Multiple routes of infection could be the cause of the high virulence and mortality rates detected in shrimp species. In particular, Penaeus vannamei, differences in isolate virulence have been observed, along with controversy over whether deletions or insertions are associated with gain or loss of virulence.The pathogenicity of three isolates from three locations in Mexico (two from Sinaloa: 'CIAD', and 'Angostura', and one from Sonora: 'Sonora') was evaluated in vivo in white shrimp (P. vannamei) infection assays. Differences were observed in the mortality rate of shrimp among the three isolates, with the Sonora isolate being the most virulent. Subsequently, the complete WSSV genomes were sequenced in depth from the tissues of infected shrimp, and assembled in reference to the genome of isolate CN01 (KT995472), identifying genome sizes for Angostura and Sonora of 289,350 bp and 288,995 bp, respectively. Where three deletion zones were identified compared to CN01 comprising 15 genes, including three envelope proteins VP41A, VP52A and VP41B, one non-structural protein ICP35 and 11 others encoding proteins whose function is currently unknown. In addition, five genes (wsv129, wsv178, wsv204, wsv249 and wsv497) show a modified number of repeat motifs. The main implications and possible effects on viral infection of these modifications are discussed.

In silico studies on the interaction of phage displayed biorecognition element (TFQAFDLSPFPS) with the structural protein VP28 of white spot syndrome virus

2020 ◽  
Vol 26 (10) ◽  
Author(s):  
Snehal Jamalpure ◽  
Gauri Panditrao ◽  
Prabir Kumar Kulabhusan ◽  
A. S. Sahul Hameed ◽  
K. M. Paknikar ◽  
...  
Keyword(s):  
In Silico ◽  
White Spot Syndrome Virus ◽  
White Spot ◽  
Structural Protein ◽  
In Silico Studies ◽  
White Spot Syndrome

scholarly journals Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship

2007 ◽  
Vol 81 (12) ◽  
pp. 6709-6717 ◽  
Author(s):  
Xuhua Tang ◽  
Jinlu Wu ◽  
J. Sivaraman ◽  
Choy Leong Hew
Keyword(s):  
Viral Infection ◽  
White Spot Syndrome Virus ◽  
Evolutionary Relationship ◽  
Viral Proteins ◽  
Envelope Proteins ◽  
Viral Envelope ◽  
White Spot ◽  
Structural Proteins ◽  
White Spot Syndrome ◽  
Effective Transfer

ABSTRACT White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 Å, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelope proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt β-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core β-barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV.

scholarly journals Polycistronic mRNAs and internal ribosome entry site elements (IRES) are widely used by white spot syndrome virus (WSSV) structural protein genes

Virology ◽  
2009 ◽  
Vol 387 (2) ◽  
pp. 353-363 ◽  
Author(s):  
Shih-Ting Kang ◽  
Jiann-Horng Leu ◽  
Han-Ching Wang ◽  
Li-Li Chen ◽  
Guang-Hsiung Kou ◽  
...  
Keyword(s):  
Internal Ribosome Entry Site ◽  
White Spot Syndrome Virus ◽  
White Spot ◽  
Structural Protein ◽  
Entry Site ◽  
Internal Ribosome Entry ◽  
White Spot Syndrome

scholarly journals RING-H2 Protein WSSV249 from White Spot Syndrome Virus Sequesters a Shrimp Ubiquitin-Conjugating Enzyme, PvUbc, for Viral Pathogenesis

2005 ◽  
Vol 79 (14) ◽  
pp. 8764-8772 ◽  
Author(s):  
Zhilong Wang ◽  
Hui Kheng Chua ◽  
Ayu A. R. A. Gusti ◽  
Fang He ◽  
Beau Fenner ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
Expression Patterns ◽  
Viral Pathogenesis ◽  
White Spot ◽  
Cellular Processes ◽  
Low Degree ◽  
Ubiquitin Conjugating Enzyme ◽  
White Spot Syndrome ◽  
Conjugating Enzyme

ABSTRACT Modification of proteins by ubiquitin is essential for numerous cellular processes. The RING-H2 finger motif has been implicated in ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Four proteins, WSSV199, WSSV222, WSSV249, and WSSV403, from white spot syndrome virus (WSSV) contain the RING-H2 motif. Here we report that WSSV249 physically interacts with a shrimp ubiquitin-conjugating enzyme, PvUbc, and mediates ubiquitination through its RING-H2 motif in the presence of E1 and PvUbc. Mutations of the putative zinc coordination residues in the RING-H2 domain of WSSV249, however, ablate ubiquitination efficiency. In addition, the RING-H2 domain of WSSV249 is capable of ubiquitination with UbcH1, UbcH2, UbcH5a, UbcH5b, UbcH5c, UbcH6, and UbcH10, respectively, exhibiting a low degree of E2 specificity. Significantly, the expression of WSSV249 and PvUbc increased during infection, as revealed by real-time PCR. Furthermore, in situ hybridization showed that WSSV249 and PvUbc display similar expression patterns in infected shrimps, and immunofluorescence and immunohistochemistry assays showed an increase of PvUbc in infected shrimp cells. These results suggest that the RING-H2 protein WSSV249 from WSSV may function as an E3 ligase via sequestration of PvUbc for viral pathogenesis in shrimp.

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