Construction and Application of a Protein Interaction Map for White Spot Syndrome Virus (WSSV)

Sangsuriya, Pakkakul; Huang, Jiun Yan; Chu, Yu Fei; Phiwsaiya, Kornsunee; Leekitcharoenphon, Pimlapas; Meemetta, Watcharachai; Senapin, Saengchan; Huang, Wei Pang; Withyachumnarnkul, Boonsirm; Flegel, Timothy W.; Lo, Chu Fang

doi:10.1074/mcp.m113.029199

Cited by 32 publications

(34 citation statements)

References 74 publications

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“…For the yeast two-hybrid screening, the PmPPAE2/pGBKT7 construct obtained above was used as the bait to screen for interacting proteins (prey) from a WSSV DNA library fused with the AD domain of pGADT7 (34). Positive interactions were indicated by growth on the high stringency media lacking leucine, tryptophan, adenine, and histidine (SD/ϪL/ϪW/ϪA/ ϪH) and by a blue color change caused by 5-bromo-4-chloro-3-indolyl-␣-D-galactoside (X-␣-gal; Apollo Scientific) in the medium.…”

Section: Journal Of Biological Chemistrymentioning

confidence: 99%

Suppression of Shrimp Melanization during White Spot Syndrome Virus Infection

Sutthangkul

Amparyup

Charoensapsri

et al. 2015

Journal of Biological Chemistry

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Background: Melanization plays a major role in invertebrate defense. Results: Suppression of shrimp melanization increased the WSSV susceptibility. The viral protein, WSSV453, interferes the proPO system via PmPPAE2 inhibition. Conclusion: Shrimp melanization has an antiviral role. WSSV overcomes this by suppressing the host proPO proteinase cascade. Significance: The regulation of shrimp melanization during WSSV infection was first demonstrated.

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Section: Journal Of Biological Chemistrymentioning

confidence: 99%

Suppression of Shrimp Melanization during White Spot Syndrome Virus Infection

Sutthangkul

Amparyup

Charoensapsri

et al. 2015

Journal of Biological Chemistry

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“…A shotgun proteomic study of WSSV (China isolate) revealed that WSSV186 (WSV131) and WSSV189 (WSV134) are WSSV structural proteins . Recently, WSSV395 and WSSV471 were identified as hub proteins interacting with over 20 WSSV proteins (Sangsuriya et al, 2014). Previously, WSSV395 (WSV339), or VP39B, was determined by Western blot and IEM analyses to be an envelope protein (Tsai et al, 2004), whilst here WSSV189 was identified as being located at the envelope of the WSSV virion by the same approach and so a likely structural protein.…”

Section: Discussionmentioning

confidence: 97%

“…Saccharomyces cerevisiae AH109 was co-transformed with pGBKT7-ALFPm3 (bait) and the prey pGADT7-WSSV ORF library (Sangsuriya et al, 2014), and transformants were selected on a synthetic defined (SD) medium lacking leucine and tryptophan (SD/-Leu/-Trp). Interaction between the bait and prey fusion proteins was indicated by the growth and blue color change on the double dropout SD/-Leu/-Trp media containing X-alpha-Gal (DDO/X).…”

Section: Yeast Two-hybrid Screeningmentioning

confidence: 99%

Anti-lipopolysaccharide factor isoform 3 from Penaeus monodon (ALFPm3) exhibits antiviral activity by interacting with WSSV structural proteins

et al. 2014

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In innate immunity, antimicrobial peptides (AMPs) play a vital role in combating microbial pathogens. Among the AMPs identified in Penaeus monodon, only anti-lipopolysaccharide factor isoform 3 (ALFPm3) has been reported to exhibit activity against white spot syndrome virus (WSSV). However, the mechanism(s) involved are still not clear. In the present study, ALFPm3-interacting proteins were screened for from a WSSV library using the yeast two-hybrid screening system, revealing the five potential ALFPm3-interacting proteins of WSSV186, WSSV189, WSSV395, WSSV458 and WSSV471. Temporal transcriptional analysis in WSSV-infected P. monodon revealed that all five of these WSSV gene transcripts were expressed in the late phase of infection (24h and 48h post-infection). Of these, WSSV189 that was previously identified as a structural protein, was selected for further analysis and was shown to be an enveloped protein by Western blot and immunoelectron microscopy analyses. The in vitro pull-down assay using recombinant WSSV189 (rWSSV189) protein as bait confirmed the interaction between ALFPm3 and WSSV189 proteins. Moreover, pre-incubation of rWSSV189 protein with rALFPm3 protein interfered with the latter's neutralization effect on WSSV in vivo, as shown by the increased cumulative mortality of shrimp injected with WSSV following prior treatment with pre-incubated rWSSV189 and rALFPm3 proteins compared to that in shrimp pre-treated with rALFPm3 protein. Thus, ALFPm3 likely performs its anti-WSSV action by binding to the envelope protein WSSV189 and possibly other WSSV structural proteins.

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“…To prepare the bait plasmid, the DNA fragment encoding L. vannamei ferritin (Lvferritin) was amplified by PCR using the specific Lvferritin-F-NdeI/Lvferritin-R-BamHI primer set listed in Table 1. The PCR products were cloned into pGBKT7-1, which is a modified pGBKT7-Rec vector in which the multiple-cloning-site sequence has been replaced by the sequence of pGADT7-Rec (23), together with the GAL4 DNA binding domain (BD). The resulting plasmid, Lvferritin-pGBKT7-1, was then transformed into Saccharomyces cerevisiae strain Y187 (Clontech) to express Lvferritin tagged with c-myc and fused with the BD domain (c-myc-Lvferritin-BD).…”

Section: Methodsmentioning

confidence: 99%

White Spot Syndrome Virus Protein Kinase 1 Defeats the Host Cell's Iron-Withholding Defense Mechanism by Interacting with Host Ferritin

Lin

Lee

Wang

et al. 2015

J Virol

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We show here that white spot syndrome virus (WSSV) ensures the availability of iron by using a previously unreported mechanism to defeat the host cell's iron-withholding defense mechanism. This defense is often implemented by ferritin, which can bind up to 4,500 iron atoms and acts to sequester free iron within the cell. WSSV's novel counterstrategy is mediated by a direct protein-protein interaction between viral protein kinase 1 (PK1) and host ferritin. PK1 interacts with both ferritin and apoferritin, suppresses apoferritin's ability to sequester free iron ions, and maintains the intracellular labile iron pool (LIP), and thus the availability of free iron is increased within cells.

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Construction and Application of a Protein Interaction Map for White Spot Syndrome Virus (WSSV)

Cited by 32 publications

References 74 publications

Suppression of Shrimp Melanization during White Spot Syndrome Virus Infection

Suppression of Shrimp Melanization during White Spot Syndrome Virus Infection

Anti-lipopolysaccharide factor isoform 3 from Penaeus monodon (ALFPm3) exhibits antiviral activity by interacting with WSSV structural proteins

White Spot Syndrome Virus Protein Kinase 1 Defeats the Host Cell's Iron-Withholding Defense Mechanism by Interacting with Host Ferritin

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