title: Molecular characterization and expression analysis of complement component C9 gene in the whitespotted bambooshark, Chiloscyllium plagiosum.
abstract: Complement system is known as highly sophisticated immune defense mechanism for antigen recognition as well as effector functions. Activation of the terminal pathway of the complement system leads to the assembly of terminal complement complexes (C5b-9), which induces the characteristic complement-mediated cytolysis. The lytic activity of shark complement involves functional analogues of mammalian C8 and C9. In this article, a full-length cDNA of C9 (CpC9) is identified from cartilaginous species, the whitespotted bambooshark, Chiloscyllium plagiosum by RACE. The CpC9 cDNA is 2263 bp in length, encoding a protein of 603 amino acids, which shares 42% and 43% identity with human and Xenopus C9 respectively. Through sequence alignment and comparative analysis, the CpC9 protein was found well conserved, with the typical modular architecture in TCCs and nearly unanimous cysteine composition from fish to mammal. Phylogenetic analysis places it in a clade with C9 orthologs in higher vertebrate and as a sister taxa to the Xenopus. Expression analysis revealed that CpC9 is constitutively highly expressed in shark liver, with much less or even undetectable expression in other tissues; demonstrating liver is the primary tissue for C9synthesis. To sum up, the structural conservation and distinctive phylogenetics might indicate the potentially vital role of CpC9 in shark immune response, though it remains to be confirmed by further study.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, No. 22 Hankou Rd, Gulou District, Nanjing 210093, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Shenshen
authors.initials: S
authors.lastname: Xu
authors.forename: Yuhao
authors.initials: Y
authors.lastname: Su
authors.forename: Boping
authors.initials: B
authors.lastname: Ye
authors.forename: Zichun
authors.initials: Z
authors.lastname: Hua
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003186:Complement C9 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012754:Sharks / Q000235:genetics* / Q000378:metabolism
chemical_list: D003186:Complement C9; D018076:DNA, Complementary; D029941:Fish Proteins; D012333:RNA, Messenger
keywords: Cartilaginous fish; Chiloscyllium plagiosum; Complement C9; CpC9; EGFP; GSP; LD PCR; LDL-R; MAC; MAC/perforin domain; MACPF; RACE; Shark complement; TCCs; TSP; Terminal complement component; UTRs; epidermal growth factor precursor domain; gene-specific primers; long distance PCR; low-density lipoprotein receptor domain; qRT-PCR; rapid amplification of cDNA ends; real-time quantitative RT-PCR; the complement component C9 for Chiloscyllium plagiosum; the membrane attack complex; the terminal complement components; thrombospondin domain; untranslated regions