PDI family thioredoxin from disk abalone (Haliotis discus discus): Responses to stimulants (PAMPs, bacteria, and viral) and functional characterization. Issue 120 (January 2022)

• Record Type: Journal Article
• Authors: Liyanage, D.S.; Omeka, W.K.M.; Sandamalika, W.M. Gayashani; Udayantha, H.M.V.; Jeong, Taehyug; Lee, Sukkyoung; Lee, Jehee
• Is Part Of: Fish & shellfish immunology. Issue 120(2022)
• Page Start: 261
• Page End: 270
• Publication Date: 2022-01
• Subjects: Antioxidant -- Haliotis discus discus -- Immune stimulation -- Oxidative stress -- Thioredoxin domain-containing 5; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2021.11.037
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Thioredoxin, a highly conserved class of proteins involved in redox signaling, is found in a range of organisms from bacteria to higher-level eukaryotes. Thioredoxin acts as an active regulatory enzyme to eliminate excessive reactive oxygen species, thereby preventing cellular damage. In this study, the cDNA sequence of thioredoxin domain-containing 5 ( AbTXNDC5 ) from the disk abalone transcriptomic database was characterized. An in silico analysis of AbTXNDC5 was performed, and its spatial and temporal expression patterns in hemocytes and gills in response to bacteria ( Vibrio parahaemolyticus, Listeria monocytogenes ), viral hemorrhagic septicemia virus, and pathogen-associated molecular pattern molecules were observed. Furthermore, AbTXNDC5 expression was examined in different developmental stages. Functional assays to explore insulin disulfide reduction, anti-apoptotic activity, and protection against hypoxic cell death of AbTXNDC5 were conducted through recombinant proteins or overexpression in cells. AbTXNDC5 contains a 1179-bp open reading frame coding for 392 amino acids. Conserved thiol-disulfide cysteine residues within two Cys-X-X-Cys motifs were found in AbTXNDC5. Quantitative real-time polymerase chain reaction indicated that healthy digestive tract and hemocyte tissues expressed high levels of AbTXNDC5 mRNA, which may protect the host from invading pathogens. Immune-challenged abalone hemocytes and gills exhibited upregulated expression of AbTXNDC5 at different time points. rAbTXNDC5 also exhibited a functional insulin disulfide reductase activity. AbTXNDC5 conferred protection to cultured cells from apoptosis and hypoxia-induced stress, compared to the pcDNA3.1 (+) transfected control cells. Therefore, AbTXNDC5 can be considered an important gene in abalones in relation to the primary immune system and regulation of redox homeostasis and confers protection from stress. Highlights: AbTXNDC5 is an important gene in redox regulation and innate immunity. AbTXNDC5 upregulated in hemocytes and gills in response to PAMPs, bacteria, and virus. RAbTXNDC5 has actively precipitated the insulin by breaking A and B chain bonds. AbTXNDC5 transfected FHM cells exhibited cell protection against apoptosis and hypoxia.