원문정보
초록
영어
β-catenin is a structural protein that makes the cell-cell connection in adherence junctions. Besides the structural functions, it also plays a role as a central transducer of the canonical Wnt signaling cascade, regulating nearly four hundred genes related to various cellular processes. Recently the immune functions of β-catenin during pathogenic invasion have gained more attention. In the present study, we elucidated the immune function of fish β-catenin by identifying and characterizing the β-catenin homolog (PhCatβ) from redlip mullet, Planiliza haematocheila. The complete open reading frame of PhCatβ consists of 2352 bp, which encodes a putative β-catenin homolog (molecular weight: 85.7 kDa). Multiple sequence alignment analysis revealed that β-catenin is highly conserved in vertebrates. Phylogenetic reconstruction demonstrated the close evolutionary relationship between PhCatβ and other fish β-catenin counterparts. The tissue distribution analysis showed the highest mRNA expression of PhCatβ in heart tissues of the redlip mullet under normal physiological conditions. While in response to pathogenic stress, the PhCatβ transcription level was dramatically increased in the spleen and gill tissues. The overexpression of PhCatβ stimulated M2 polarization and cell proliferation of murine RAW 264.7 macrophage. In fish cells, the overexpression of PhCatβ resulted in a significant upregulation of antiviral gene transcription and vice versa. Moreover, the overexpression of PhCatβ could inhibit the replication of VHSV in FHM cells. Our results strongly suggest that PhCatβ plays a role in macrophage activation and antiviral immune response in redlip mullet.
목차
1. Introduction
2. Materials and methods
2.1. Rearing of fish, tissue collection, and transcriptomic database construction
2.2. Bioinformatics analysis
2.3. Tissue collection, mRNA extraction, and gene expression analysis from tissues of healthy and challenged fish
2.4. Analysis of spatial and temporal expression of PhCatβ by qPCR
2.5. Plasmid construction for the overexpression of PhCatβ
2.6. Subcellular localization of PhCatβ in FHM cells
2.7. Macrophage polarization and proliferation activities of PhCatβ
2.8. Analysis of antiviral gene transcription after overexpression or knockdown of PhCatβ
2.9. Antiviral activity assay of PhCatβ against VHSV
3. Results
3.1. Sequence and phylogeny analysis of PhCatβ
3.2. Tissue-specific transcription of PhCatβ and temporal transcription response to pathogenic stimuli
3.3. Subcellular localization of PhCatβ in FHM cells
3.4. Macrophage M2 polarization and cell proliferation effect of PhCatβ overexpression
3.5. Antiviral functions of PhCatβ
4. Discussion
5. Conclusion
References