Identification and characterization of syntenin binding protein in the black tiger shrimp Penaeus monodon.
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 (alpha2M). A GST pull-down assay showed that GST-alpha2M, 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 alpha2M. 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 alpha2M. Furthermore, WSSV-infected Pm showed a significant upregulation of the alpha2M message, suggesting that both syntenin and its protein partner alpha2M are upregulated in the acute phase of a WSSV infection. Taken together with a previous report showing the co-localization of alpha2M and syntenin in the exosome of a dendritic cell line, it is likely that syntenin, through its interaction with alpha2M, plays an important role in the immune defense mechanisms of viral infections of shrimps.
Tonganunt
Moltira
M
Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand.
Phongdara
Amornrat
A
Chotigeat
Wilaiwan
W
Fujise
Kenichi
K
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004266:DNA Virus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology; D015870:Gene Expression; D006434:Hemocytes / Q000378:metabolism; D033561:Penaeidae / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D046848:White spot syndrome virus 1 / Q000472:pathogenicity; D000511:alpha-Macroglobulins / Q000235:genetics / Q000378:metabolism
D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; D000511:alpha-Macroglobulins; D004247:DNA