title: Two carboxypeptidase counterparts from rock bream (Oplegnathus fasciatus): molecular characterization, genomic arrangement and immune responses upon pathogenic stresses.
abstract: Carboxypeptidases (CPs) are proteases that hydrolyze C-terminal peptide bonds. They are involved in regulating the complement system of the immune system. Here, we report the molecular characterization and immune response of two carboxypeptidases, named carboxypeptidase A (Rb-CPA) and carboxypeptidase N1 (Rb-CPN1), from rock bream. The genomic sequence of Rb-CPA contains 12 exons interrupted by 11 introns, while the genomic sequence of Rb-CPN1 has 9 exons and 8 introns. The cDNA sequence of Rb-CPA encodes a 421-amino-acid (AA) polypeptide (48kDa), and the cDNA of Rb-CPN1 encodes a 448-AA polypeptide (51kDa). The amino acid sequences of Rb-CPA and Rb-CPN1 were found to harbor two characteristic Zn-binding signature domains and a peptidase-M14 Zn carboxypeptidase site. Pairwise analysis revealed that Rb-CPA and Rb-CPN1 had the highest identity with the corresponding proteins from Anoplopoma fimbria (87.6%) and Dicentrarchus labrax (96.9%), respectively. qPCR results indicated that Rb-CPA and Rb-CPN1 were constitutively expressed mainly in the kidney, heart, liver, and head kidney. Both genes were transcriptionally regulated in the liver upon challenge with pathogenic bacteria (Streptococcus iniae, Edwardsiella tarda), rock bream iridovirus (RBIV), and the immune modulators polyinosinic:polycytidylic acid and lipopolysaccharide. Taken together, our findings suggest that Rb-CPA and Rb-CPN1 have immune-related functions in rock bream.
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: G I
authors.initials: GI
authors.lastname: Godahewa
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: W D Niroshana
authors.initials: WD
authors.lastname: Wickramaarachchi
authors.affiliation: Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: Ilson
authors.initials: I
authors.lastname: Whang
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: S D N K
authors.initials: SD
authors.lastname: Bathige
authors.affiliation: Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors.forename: Bong-Soo
authors.initials: BS
authors.lastname: Lim
authors.affiliation: Division of Marine Environment and Bioscience, Korea Maritime University, Busan 606-791, Republic of Korea.
authors.forename: Cheol Young
authors.initials: CY
authors.lastname: Choi
authors.affiliation: Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; College of Veterinary Medicine, Chungnam National University, Yuseong-gu, Daejeon 305-764, Republic of Korea. Electronic address: mahanama@cnu.ac.kr.
authors.forename: Mahanama
authors.initials: M
authors.lastname: De Zoysa
authors.affiliation: Genetics & Breeding Research Center, National Fisheries Research & Development Institute, Geoje 656-842, Republic of Korea.
authors.forename: Jae Koo
authors.initials: JK
authors.lastname: Noh
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea. Electronic address: jehee@jejunu.ac.kr.
authors.forename: Jehee
authors.initials: J
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002268:Carboxypeptidases / Q000235:genetics / Q000276:immunology*; D020609:Edwardsiella tarda / Q000276:immunology*; D004756:Enterobacteriaceae Infections / Q000201:enzymology / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D005393:Fish Diseases / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D008969:Molecular Sequence Data; D010473:Perciformes; D010802:Phylogeny; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013291:Streptococcus / Q000276:immunology*
chemical_list: D012329:RNA, Bacterial; D002268:Carboxypeptidases
keywords: Anaphylatoxin; Carboxypeptidases; Genome; Immune challenge; Rock bream