Identification and characterization of protein 14-3-3 in carcinogenic liver fluke Opisthorchis viverrini
Alok Kafle1, Pranom Puchadapirom2, Sirikanya Plumworasawat1
1Department of Microbiology, Faculty of Science, Mahidol University, 272 Rama 6 Rd., Phyathai, Rachthewee, Bangkok 10400, Thailand.
Abstract:
Protein 14-3-3s are abundant phospho-serine/threonine binding proteins, which are highly conserved among eukaryotes. Members of this protein family mediate metabolism and signal transduction networks through binding to hundreds of other protein partners. Protein 14-3-3s have been studied in other species of parasitic helminthes, but little is known about this protein in the carcinogenic liver fluke Opisthorchis viverrini. In this study, we identified and characterized protein 14-3-3s of O. viverrini. Seven protein 14-3-3 encoded sequences were retrieved from the O. viverrini genome database. Multiple alignment and phylogenetic analysis were performed. Two isoforms (protein 14-3-3 zeta and protein 14-3-3 epsilon) that have been previously found in the excretory-secretory (ES) products of O. viverrini were produced as recombinant protein in E. coli and the proteins were then used to immunize mice to obtain specific antibodies. Western blot analysis showed that both proteins were detected in all obtainable developmental stages of O. viverrini and the ES products. Immunolocalization revealed that both isoforms were expressed throughout tissues and organs except the gut epithelium. The highest expression was observed in testes especially in developing spermatocytes, suggesting their role in spermatogenesis. Prominent expression was also detected on tegumental surface of the parasite and on epical surface of bile duct epithelium indicates their additional role in host-parasite interaction. These findings indicate that protein 14-3-3s play important role in the life cycle of the parasite and might be involved in the pathogenesis of O. viverrini infection.
Insights
This study identified and characterized 14-3-3 proteins in the liver fluke Opisthorchis viverrini. These proteins are present in all life stages and may play roles in parasite development and host interaction.
Area of Science:
- Parasitology
- Molecular Biology
- Biochemistry
Background:
- 14-3-3 proteins are conserved eukaryotic proteins involved in diverse cellular processes.
- Their role in the carcinogenic liver fluke Opisthorchis viverrini remains largely uncharacterized.
- Understanding these proteins is crucial for insights into parasite biology and pathogenesis.
Purpose of the Study:
- To identify and characterize the 14-3-3 protein family in Opisthorchis viverrini.
- To investigate the expression patterns and potential functions of O. viverrini 14-3-3 isoforms.
- To explore the involvement of these proteins in parasite development and host-parasite interactions.
Main Methods:
- Bioinformatic analysis of O. viverrini genome database for 14-3-3 sequences.
- Phylogenetic analysis and multiple sequence alignment.
- Recombinant protein expression (14-3-3 zeta and epsilon) in E. coli.
- Antibody production and Western blot analysis across developmental stages and excretory-secretory products.
- Immunolocalization studies to determine tissue-specific expression.
Main Results:
- Seven 14-3-3 encoding sequences were identified in the O. viverrini genome.
- Two isoforms, 14-3-3 zeta and epsilon, were successfully expressed and detected in all parasite life stages and excretory-secretory products.
- Immunolocalization revealed expression in various tissues, notably high in testes (spermatogenesis), on the parasite tegument, and bile duct epithelium.
- Expression patterns suggest roles in spermatogenesis, host-parasite interaction, and potentially pathogenesis.
Conclusions:
- 14-3-3 proteins are integral components of Opisthorchis viverrini.
- These proteins are likely involved in critical life cycle processes, including reproduction and host interaction.
- Further research into O. viverrini 14-3-3 proteins may reveal targets for therapeutic intervention against opisthorchiasis.
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