Identification and characterization of syntenin binding protein in the black tiger shrimp Penaeus monodon

Moltira Tonganunt, Amornrat Phongdara, Wilaiwan Chotigeat, Kenichi Fujise

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Shrimp exhibit a diverse response to viral infection that is manifested in drastic up- and down-regulations of a variety of genes. In our previous work, we identified syntenin of the shrimp Penaeus monodon (Pm) as a dynamic responder to white spot syndrome virus (WSSV) infection, its message being greatly upregulated in the acute phase of the infection. In order to further explore the link between Pm-syntenin and viral infection, we performed a yeast two-hybrid screening of a P. monodon cDNA library, using Pm-syntenin as bait. One of the molecules that specifically interacted with Pm-syntenin was the receptor-binding domain of alpha-2-macroglobulin (α2M). A GST pull-down assay showed that GST-α2M, but not GST alone, was capable of co-precipitating syntenin. Another GST pull-down assay showed that GST-syntenin, but not GST alone, was capable of co-precipitating α2M. In addition, mutant analyses showed that the N-terminal 131 amino acids of syntenin were both necessary and sufficient to bind the C-terminus receptor-binding domain of α2M. Furthermore, WSSV-infected Pm showed a significant upregulation of the α2M message, suggesting that both syntenin and its protein partner α2M are upregulated in the acute phase of a WSSV infection. Taken together with a previous report showing the co-localization of α2M and syntenin in the exosome of a dendritic cell line, it is likely that syntenin, through its interaction with α2M, plays an important role in the immune defense mechanisms of viral infections of shrimps.

Original languageEnglish (US)
Pages (from-to)135-145
Number of pages11
JournalJournal of Biotechnology
Volume120
Issue number2
DOIs
StatePublished - Nov 4 2005
Externally publishedYes

Fingerprint

Syntenins
Penaeidae
Tigers
Viruses
Macroglobulins
Carrier Proteins
Assays
Virus Diseases
White spot syndrome virus 1
Yeast
Amino acids
Screening
Genes
Proteins
Molecules
Up-Regulation
Exosomes
alpha-Macroglobulins
Gene Library
Dendritic Cells

Keywords

  • Alpha-2-macroglobulin
  • Immune
  • Syntenin
  • White spot syndrome virus
  • Yeast two-hybrid

ASJC Scopus subject areas

  • Biotechnology

Cite this

Identification and characterization of syntenin binding protein in the black tiger shrimp Penaeus monodon. / Tonganunt, Moltira; Phongdara, Amornrat; Chotigeat, Wilaiwan; Fujise, Kenichi.

In: Journal of Biotechnology, Vol. 120, No. 2, 04.11.2005, p. 135-145.

Research output: Contribution to journalArticle

Tonganunt, Moltira ; Phongdara, Amornrat ; Chotigeat, Wilaiwan ; Fujise, Kenichi. / Identification and characterization of syntenin binding protein in the black tiger shrimp Penaeus monodon. In: Journal of Biotechnology. 2005 ; Vol. 120, No. 2. pp. 135-145.
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abstract = "Shrimp exhibit a diverse response to viral infection that is manifested in drastic up- and down-regulations of a variety of genes. In our previous work, we identified syntenin of the shrimp Penaeus monodon (Pm) as a dynamic responder to white spot syndrome virus (WSSV) infection, its message being greatly upregulated in the acute phase of the infection. In order to further explore the link between Pm-syntenin and viral infection, we performed a yeast two-hybrid screening of a P. monodon cDNA library, using Pm-syntenin as bait. One of the molecules that specifically interacted with Pm-syntenin was the receptor-binding domain of alpha-2-macroglobulin (α2M). A GST pull-down assay showed that GST-α2M, but not GST alone, was capable of co-precipitating syntenin. Another GST pull-down assay showed that GST-syntenin, but not GST alone, was capable of co-precipitating α2M. In addition, mutant analyses showed that the N-terminal 131 amino acids of syntenin were both necessary and sufficient to bind the C-terminus receptor-binding domain of α2M. Furthermore, WSSV-infected Pm showed a significant upregulation of the α2M message, suggesting that both syntenin and its protein partner α2M are upregulated in the acute phase of a WSSV infection. Taken together with a previous report showing the co-localization of α2M and syntenin in the exosome of a dendritic cell line, it is likely that syntenin, through its interaction with α2M, plays an important role in the immune defense mechanisms of viral infections of shrimps.",
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