1. Molecular characterization and immune regulatory, antioxidant, and antiapoptotic activities of thioredoxin domain-containing protein 17 (TXNDC17) in yellowtail clownfish (Amphiprion clarkii)
- Author
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Sukkyoung Lee, W.M. Gayashani Sandamalika, Chaehyeon Lim, Hyerim Yang, Anushka Vidurangi Samaraweera, Jehee Lee, Kishanthini Nadarajapillai, and H.M.V. Udayantha
- Subjects
Fish Proteins ,Lipopolysaccharides ,0301 basic medicine ,Aquatic Science ,Fish Diseases ,03 medical and health sciences ,Thioredoxins ,Complementary DNA ,Gene expression ,Animals ,Environmental Chemistry ,Amino Acid Sequence ,Phylogeny ,Vibrio ,chemistry.chemical_classification ,Innate immune system ,biology ,Molecular mass ,Vibrio harveyi ,Gene Expression Profiling ,Cytochrome c ,Fishes ,04 agricultural and veterinary sciences ,General Medicine ,biology.organism_classification ,Immunity, Innate ,Amino acid ,Poly I-C ,030104 developmental biology ,Gene Expression Regulation ,Biochemistry ,chemistry ,Vibrio Infections ,040102 fisheries ,biology.protein ,0401 agriculture, forestry, and fisheries ,Thioredoxin ,Sequence Alignment - Abstract
Thioredoxin domain-containing protein 17 (TXNDC17) is an important, highly conserved oxidoreductase protein, ubiquitously expressed in all living organisms. It is a small (~14 kDa) protein mostly co-expressed with thioredoxin 1 (TRx1). In the present study, we obtained the TXNDC17 gene sequence from a previously constructed yellowtail clownfish (Amphiprion clarkii) (AcTXNDC17) database and studied its phylogeny as well as the protein's molecular characteristics, antioxidant, and antiapoptotic effects. The full length of the AcTXNDC17 cDNA sequence was 862 bp with a 372 bp region encoding a 123 amino acid (aa) protein. The predicted molecular mass and isoelectric point of AcTXNDC17 were 14.2 kDa and 5.75, respectively. AcTXNDC17 contained a TRX-related protein 14 domain and a highly conserved N-terminal Cys43-Pro44-Asp45-Cys46 motif. qPCR analysis revealed that AcTXNDC17 transcripts were ubiquitously and differently expressed in all the examined tissues. AcTXNDC17 expression in the spleen tissue was significantly upregulated in a time-dependent manner upon stimulation with lipopolysaccharide (LPS), polyinosinic-polycytidylic (poly I:C), and Vibrio harveyi. Besides, LPS-induced intrinsic apoptotic pathway (TNF-α, caspase-8, Bid, cytochrome C, caspase-9, and caspase-3) gene expression was significantly lower in AcTXNDC17-overexpressing RAW264.7 cells, as were NF-κB activation and nitric oxide (NO) production. Furthermore, the viability of H2O2-stimulated macrophages was significantly improved under AcTXNDC17 overexpression. Collectively, our findings indicate that AcTXNDC17 is involved in the innate immune response of the yellowtail clownfish.
- Published
- 2021