scholarly journals Hijacking of Host Calreticulin is Required for the White Spot Syndrome Virus Replication Cycle.

2014 ◽  
pp. JVI.01014-14 ◽  
Author(s):  
Apiruck Watthanasurorot ◽  
Enen Guo ◽  
Sirinit Tharntada ◽  
Chu-Fang Lo ◽  
Kenneth Söderhäll ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
White Spot Syndrome Virus ◽  
Replication Cycle ◽  
Host Protein ◽  
Host Cells ◽  
White Spot ◽  
Structural Proteins ◽  
Dna Virus ◽  
White Spot Syndrome ◽  
Viral Structural Proteins

We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP28 with CRT does not promote WSSV entry, and CRT-VP15 interaction was detected in the viral genome in virally infected host cells, and thus may have an effect on WSSV replication. Moreover, CRT was detected in the viral envelope of purified WSSV virions. CRT was also found to be of high importance for proper oligomerization of the viral structural proteins VP26 and VP28, and when CRT glycosylation was blocked with tunicamycin, a significant decrease in both viral replication and assembly was detected. Together, these findings suggest that CRT confers several advantages to WSSV from the initial steps of WSSV infection, to the assembly of virions. Therefore, CRT is required as a “vital factor” and is hijacked by WSSV for its replication cycle.ImportanceWhite spot syndrome virus (WSSV) is a double-stranded DNA virus and the cause of a serious disease in a wide range of crustaceans that often leads to high mortality rates. We have previously shown that the protein calreticulin (CRT), which resides in the endoplasmic reticulum (ER) of the cell, is important in the host response to the virus. In this report, we show that the virus uses this host protein to enter the cell and to make the host produce new viral structural proteins. Through its interaction with two viral proteins, the virus “hijacks” host calreticulin and uses it for its own needs. These findings provide new insight into the interaction between a large DNA virus and the host protein CRT, and may help in understanding the viral infection process in general.

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 Identification of a Novel Nonstructural Protein, VP9, from White Spot Syndrome Virus: Its Structure Reveals a Ferredoxin Fold with Specific Metal Binding Sites

2006 ◽  
Vol 80 (21) ◽  
pp. 10419-10427 ◽  
Author(s):  
Yang Liu ◽  
Jinlu Wu ◽  
Jianxing Song ◽  
J. Sivaraman ◽  
Choy L. Hew
Keyword(s):  
Binding Sites ◽  
Metal Ion ◽  
White Spot Syndrome Virus ◽  
Nonstructural Protein ◽  
Divalent Metal ◽  
Full Length ◽  
Host Cells ◽  
White Spot ◽  
E2 Protein ◽  
White Spot Syndrome

ABSTRACT White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP9, a full-length protein of WSSV, encoded by open reading frame wsv230, was identified for the first time in the infected Penaeus monodon shrimp tissues, gill, and stomach as a novel, nonstructural protein by Western blotting, mass spectrometry, and immunoelectron microscopy. Real-time reverse transcription-PCR demonstrated that the transcription of VP9 started from the early to the late stage of WSSV infection as a major mRNA species. The structure of full-length VP9 was determined by both X-ray and nuclear magnetic resonance (NMR) techniques. It is the first structure to be reported for WSSV proteins. The crystal structure of VP9 revealed a ferredoxin fold with divalent metal ion binding sites. Cadmium sulfate was found to be essential for crystallization. The Cd2+ ions were bound between the monomer interfaces of the homodimer. Various divalent metal ions have been titrated against VP9, and their interactions were analyzed using NMR spectroscopy. The titration data indicated that VP9 binds with both Zn2+ and Cd2+. VP9 adopts a similar fold as the DNA binding domain of the papillomavirus E2 protein. Based on our present investigations, we hypothesize that VP9 might be involved in the transcriptional regulation of WSSV, a function similar to that of the E2 protein during papillomavirus infection of the host cells.

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 Characterization of an envelope protein (VP110) of White spot syndrome virus

2006 ◽  
Vol 87 (7) ◽  
pp. 1909-1915 ◽  
Author(s):  
Li Li ◽  
Shumei Lin ◽  
Feng Yang
Keyword(s):  
Mass Spectrometry ◽  
Envelope Protein ◽  
White Spot Syndrome Virus ◽  
Open Reading Frame ◽  
Host Cells ◽  
White Spot ◽  
Genome Database ◽  
Reading Frame ◽  
Rgd Motif ◽  
White Spot Syndrome

A protein of 110 kDa (termed VP110) from the envelope fraction of White spot syndrome virus (WSSV) was identified by SDS-PAGE and mass spectrometry. The resulting amino acid sequence matched an open reading frame (wsv035) containing an Arg–Gly–Asp (RGD) motif in the WSSV genome database. To validate the mass-spectrometry result, the C-terminal segment of the wsv035 open reading frame was expressed in Escherichia coli as a fusion protein, which was used to produce specific antibody. Analysis by Western blotting and immunoelectron microscopy demonstrated that VP110 was an envelope protein of WSSV. An interaction analysis was performed between VP110 and the host cells, using a fluorescence assay and a competitive-inhibition assay. The results showed that VP110 was capable of attaching to host cells and that adhesion could be inhibited by synthetic RGDT peptides, suggesting that the RGD motif in the VP110 sequence may play a role in WSSV infection.

scholarly journals Penaeidins are a novel family of antiviral effectors against WSSV in shrimp

2018 ◽  
Author(s):  
Bang Xiao ◽  
Qihui Fu ◽  
Shengwen Niu ◽  
Haoyang Li ◽  
Kai Lǚ ◽  
...  
Keyword(s):  
Viral Infection ◽  
Signaling Pathways ◽  
Outer Surface ◽  
Phagocytic Activity ◽  
White Spot Syndrome Virus ◽  
Antiviral Immunity ◽  
White Spot ◽  
Structural Proteins ◽  
First Report ◽  
White Spot Syndrome

AbstractPenaeidins are members of a family of key effectors with broad anti-bacterial activities in penaeid shrimp. However, the function of penaeidins in antiviral immunity is rarely reported and remains largely unknown. Herein, we uncovered that penaeidins are a novel family of antiviral effectors against white spot syndrome virus (WSSV). Firstly, RNAi in vivo mediated knockdown of each penaeidin from four identified penaeidins from Litopenaeus vannamei resulted in elevated viral loads and rendered shrimp more susceptible to WSSV, whilst the phenotype of survival rate in penaeidin-silenced shrimp can be rescued via the injection of recombinant penaeidin proteins. Moreover, pull-down assays demonstrated the conserved PEN domain of penaeidin was able to interact with WSSV structural proteins. Furthermore, we observed that colloidal gold-labeled penaeidins were located on the outer surface of the WSSV virion. By infection-blocking assay, we observed that hemocytes had lower viral infection rates in the group of WSSV preincubated with penaeidins than those of control group. Phagocytic activity analysis further showed that penaeidins were able to inhibit phagocytic activity of hemocytes against WSSV Taken together, these results suggest that penaeidins specifically binds to WSSV virion by interacting with its structural proteins, thus preventing viral infection that confers host against WSSV. In addition, dual-luciferase assay and EMSA assay demonstrated that penaeidins were regulated by Dorsal and Relish, two transcription factors of the canonical Toll and IMD pathway, respectively. To our best knowledge, this is the first report on uncovering the antiviral function of penaeidins in the innate immune system of shrimp.ImportancesWhite spot syndrome, caused by white spot syndrome virus (WSSV), is the most serious disease in shrimp aquaculture, which has long been a scourge of cultured shrimp industry. Herein, we provided some substantial evidences to indicate that penaeidins are a novel family of effectors with antiviral activity against WSSV in shrimp. Penaeidins such as BigPEN, PEN2 and PEN3 were able to interact with the outer surface of WSSV virion via binding to viral structural proteins, and thus preventing viral entry host cells. In addition, we demonstrated that the Toll and IMD signaling pathways can regulate the transcriptional expression of penaeidins, which may suggest an important role of the conserved innate signaling pathways in antiviral immunity. This is the first report of the antiviral mechanism of penaeidins in shrimp, which may provide some new insights into strategies to control WSSV infection in shrimp farms.

scholarly journals Protection of Caridea Against White Spot Syndrome Virus

2020 ◽  
Vol 2 (1) ◽  
pp. 61-62
Keyword(s):  
Viral Entry ◽  
White Spot Syndrome Virus ◽  
Viral Proteins ◽  
Shrimp Farming ◽  
Host Cells ◽  
White Spot ◽  
Economic Losses ◽  
Viral Pathogen ◽  
Antisense Constructs ◽  
White Spot Syndrome

White spot syndrome virus (WSSV) belongs to a new virus family, Nimaviridae, genus Whispovirus and contains a large circular double-stranded DNA genome of 292,967 bp. WSSV virions are ellipsoid to bacilliform, enveloped particles with a distinctive tail-like appendage at one end. They can be found throughout the body of infected shrimp. The virions contain one nucleocapsid with a typical striated appearance and 5 major and at least 13 minor proteins. WSSV, which was first discovered in Southeast Asia around 1992, is currently the most serious viral pathogen of shrimp worldwide. It causes up to 100% mortality within 7 to 10 days in commercial shrimp farms, resulting in large economic losses amounting to billions of US dollars across different countries to the shrimp farming industry. In a natural situation, shrimp become infected through both oral and water-borne routes, and the gills are thought to be a major point of viral entry. Considering the global economic and sociological importance of shrimp farming and its continued high growth, the development of novel control measures becomes necessary against the outbreak of WSSV. A number of strategies have been used to control WSSV, each with some limitations. Conventional control strategies such as improvement of environmental conditions, stocking of pathogen-free post-larvae, and augmentation of disease resistance by oral immune-stimulants or probiotics are currently employed to control WSSV infection. Use of recombinant viral proteins as vaccines that induce a specific immune response and protection has been demonstrated to control WSSV. Other studies have shown successful vaccination of shrimp with DNA vaccines that have prolonged effects. The RNA interference (RNAi) mediated silencing of targeted viral mRNAs holds tremendous potential for controlling shrimp diseases. The silencing of viruses using RNAi has been experimentally demonstrated for WSSV in shrimp by injecting or feeding synthetic siRNA, long double-stranded RNA (dsRNA), and short/long-hairpin RNA (shRNA/lhRNA) prepared by in vitro transcription or expressed in bacteria. In addition to targeting viral proteins, protection of WSSV has also been achieved by dsRNA targeted against shrimp PmRab7, a protein important for viral entry into the host cells. Antisense constructs offered strong protection in WSSV challenged shrimp, P. monodon, with a corresponding decrease in viral load. Antisense constructs expressing VP24 and VP28 offered the best protection with a consistent reduction in WSSV copy number in both cell culture and in experimental shrimp. The advantage of using antisense constructs is their lack of toxicity and immunogenicity and their high specificity towards the desired target. The usage of edible pellet feed coated with dsRNA against WSSV has shown promising results. Overall, the present investigation clearly demonstrates that it is possible to induce strong protection in shrimp against WSSV infection using host promoter-driven antisense constructs in controlled laboratory-scale experiments. However, it is important to develop a simple and efficient delivery system for extending this study to the field level.

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 A 3D Model of the Membrane Protein Complex Formed by the White Spot Syndrome Virus Structural Proteins

PLoS ONE ◽  
2010 ◽  
Vol 5 (5) ◽  
pp. e10718 ◽  
Author(s):  
Yun-Shiang Chang ◽  
Wang-Jing Liu ◽  
Cheng-Chung Lee ◽  
Tsung-Lu Chou ◽  
Yuan-Ting Lee ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Complex ◽  
3D Model ◽  
White Spot Syndrome Virus ◽  
White Spot ◽  
Structural Proteins ◽  
Membrane Protein Complex ◽  
White Spot Syndrome

scholarly journals Perbandingan Metode PCR Konvensional Dengan Metode PCR Portable Kit Untuk Deteksi WSSV Pada Udang Vannamei.

2021 ◽  
Vol 9 (1) ◽  
Author(s):  
R. Rara Amaliah Fitri ◽  
Farida . ◽  
Eko Prasetio
Keyword(s):  
Plasmid Dna ◽  
White Spot Syndrome Virus ◽  
White Spot ◽  
Dna Virus ◽  
White Spot Syndrome

Udang vannamei merupakan komoditi yang mendominasi pertambakan dipesisir wilayah Kalimantan Barat. Usaha budidaya tersebut tumbuh pesat dengan resiko penyebaran penyakit yang juga tinggi. Serangan penyakit yang paling umum dan sering ditemukan pada budidaya udang vannamei adalah WSSV ( White spot Syndrome Virus). Identifikasi penyakit virus WSSV di SKIPM Pontianak dilakukan dengan dua metode yang berbeda yaitu metode PCR Konvensional dan Portable Kit PCR. Metode PCR konvensional merupakan salah satu alternatif untuk deteksi penyakit virus yang cukup akurat dan relatif lebih murah, jika dibandingkan dengan metode lain yang sedang berkembang saat ini seperti metode PCR Portable Kit yang proses deteksi lebih singkat karena tidak memerlukan tahap elektroforesis. Melihat dari kelebihan dan kekurangan masing- masing metode tersebut, sehingga menimbulkan alasan untuk mengetahui lebih lanjut tentang presisi dan akurasi sensitivitas dari kedua metode tersebut dalam mendeteksi infeksi virus WSSV pada udang vannamei. Pengamatan dilakukan pada variabel plasmid kontrol (+) masing – masing kit uji, plasmid DNA sampel dan plasmid DNA virus WSSV dengan indikator hasil pemeriksaan positif, negatif, terdeteksi maupun tidak terdeteksi. Setiap hasil pemeriksaan variabel pengamatan akan diberi nilai pembobotan tertentu, untuk mengetahui presisi dan akurasi sensitivitas dari kedua metode PCR dalam mendeteksi infeksi virus WSSV pada udang vannamei.

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