Characterisation of a novel Type I crustin involved in antibacterial and antifungal responses in the red claw crayfish, Cherax quadricarinatus. (January 2016)

• Record Type: Journal Article
• Authors: Yu, Ai-Qing; Shi, Yong-Hai; Wang, Qun
• Is Part Of: Fish & shellfish immunology. Volume 48(2016:Jan.)
• Page Start: 30
• Page End: 38
• Publication Date: 2016-01
• Subjects: Crustin -- Innate immunity -- Pathogen-associated molecular patterns (PAMPs) -- Cherax quadricarinatus; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2015.11.019
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Antimicrobial peptides are important immune effectors involved in mediating innate immune responses against intruding pathogens. Here, we successfully isolated and characterized a novel Type I crustin from the red claw crayfish Cherax quadricarinatus . The full-length cDNA encoded by this gene, designated CqCrs, comprised 608 bp, containing a 5′-untranslated region (UTR) of 55 bp, a 3′-UTR of 229 bp with a poly (A) tail, and an open reading frame (ORF) of 324 bp encoding a polypeptide of 107 amino acids. The deduced amino acid sequence of Cq Crs exhibited a configuration typical of other crustacean Type I crustin orthologs, including one signal peptide region at the N-terminus between residues 1 and 16 and a long whey acidic protein (WAP) domain at the C-terminus between residues 60 and 107, along with a WAP-type "four-disulfide core" motif. Phylogenetic analysis showed that CqCrs was clustered first with other crustacean Type I crustins, then with other crustacean Type II crustins, and finally with other crustacean Type III crustins. Transcription of CqCrs was detected in all tissues, especially in immune tissues and was differentially induced in hemocytes post-stimulation with β-1, 3-glucan, lipopolysaccharides (LPS) and peptidoglycans (PG) at selected time-points. To clarify the biological activity of Cq Crs, the recombinant Cq Crs protein (r Cq Crs) was constructed and expressed in Escherichia coli BL21 (DE3). Purified r Cq Crs bound to diverse bacteria and inhibited the growth of different microbes to varying degrees. These findings suggest that Cq Crs is involved in a specific innate immune recognition and defense mechanisms against bacterial and fungal in C. quadricarinatus . Highlights: Novel crayfish crustin Cq Crs was cloned and characterized. Cq Crs is a ubiquitous protein in crayfish, especially in immune tissues. CqCrs expression in crayfish hemocytes varies after pathogenic stimulation. r Cq Crs exhibited different binding and antimicrobial activity to diverse pathogens. Cq Crs may be involved in immune responses against bacteria and fungi.