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Published on: January 20, 2017
Anti-lipopolysaccharide factor isoform 3 from Penaeus monodon (ALFPm3) exhibits antiviral activity by interacting
Sivalee Suraprasit1, Thanachai Methatham1, Phattarunda Jaree1
1Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Abstract:
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.
Insights
Antimicrobial peptides like anti-lipopolysaccharide factor isoform 3 (ALFPm3) combat pathogens. This study reveals ALFPm3 binds to WSSV envelope protein WSSV189, explaining its antiviral action against white spot syndrome virus (WSSV).
Area of Science:
- * Marine biology and immunology
- * Invertebrate pathology and disease resistance
- * Molecular virology and host-pathogen interactions
Background:
- * Antimicrobial peptides (AMPs) are crucial for innate immunity against microbial threats.
- * Anti-lipopolysaccharide factor isoform 3 (ALFPm3) is an AMP in Penaeus monodon with known activity against white spot syndrome virus (WSSV).
- * The precise mechanism of ALFPm3's antiviral activity against WSSV remains unclear.
Purpose of the Study:
- * To identify proteins interacting with ALFPm3 within the WSSV.
- * To elucidate the molecular mechanism by which ALFPm3 neutralizes WSSV.
- * To investigate the role of WSSV structural proteins in ALFPm3-mediated antiviral defense.
Main Methods:
- * Yeast two-hybrid screening to identify ALFPm3-interacting WSSV proteins.
- * Temporal transcriptional analysis of WSSV genes in infected shrimp.
- * Western blot and immunoelectron microscopy to characterize WSSV189.
- * In vitro pull-down assays to confirm protein interactions.
- * In vivo shrimp infection models to assess the impact of protein interactions on WSSV neutralization.
Main Results:
- * Five potential ALFPm3-interacting WSSV proteins were identified: WSSV186, WSSV189, WSSV395, WSSV458, and WSSV471.
- * These WSSV genes were predominantly expressed during the late phase of infection.
- * WSSV189, an enveloped structural protein, directly interacts with ALFPm3.
- * Pre-incubation of WSSV189 and ALFPm3 proteins diminished ALFPm3's WSSV neutralization efficacy in vivo, leading to increased shrimp mortality.
Conclusions:
- * ALFPm3 exerts its anti-WSSV activity by binding to the WSSV envelope protein WSSV189.
- * This interaction likely disrupts the virus's ability to infect host cells.
- * ALFPm3 may also interact with other WSSV structural proteins, contributing to its broad antiviral effect.
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