title: Cloning, identification of the two cytokine receptor family B subunits CRFB1 and CRFB5 from grass carp (Ctenopharyngodon idella).
abstract: Similar to the mammalian counterparts, fish type I interferon (IFN) performs its potential biological activities via binding to the corresponding receptor on target cell membrane. Fish type I IFN receptor, a kind of enzyme-linked receptor, consists of two subunits and belongs to the class II cytokine receptor family B (CRFB). In the present study, we cloned and identified two putative grass carp (Ctenopharyngodon idella) type I interferon receptor subunits (termed CiCRFB1 and CiCRFB5) by homology cloning techniques. Phylogenetic tree analysis suggested that CiCRFB1 and CiCRFB5 shared highly homology to Danio rerio CRFB1 and CRFB5 respectively. CiCRFB1 and CiCRFB5 were up-regulated after the stimulation with Grass Carp Hemorrhagic Virus (GCHV) and Polyinosinic-polycytidylic acid (Poly I:C), indicating that they are related to the intracellular antiviral activity. In order to know more about the roles of CiCRFB1 and CiCRFB5 in the process, the extracellular domains of CiCRFB1 (CiCRFB1-EC) and CiCRFB5 (CiCRFB5-EC), as well as grass carp type I IFN (CiIFN) were expressed in Escherichia coli BL21, and purified by affinity chromatography with the Ni-NTA His-Bind resin. Cross-linking reactions were employed to analyze the affinity of the ligand (CiIFN) with the two putative receptor subunits (CiCRFB1-EC and CiCRFB5-EC). The result suggested the formation of (CiCRFB5)2 homodimer was more easily than that of (CiCRFB1)2 under the induction of CiIFN in vitro. However, CiIFN was inclined to bind to (CiCRFB1)2 homodimer. Interestingly, although CiIFN seemed unable to facilitate the formation of (CiCRFB1 + CiCRFB5) heterodimer in the absence of DSS cross linker, however it can bind to the heterodimer in the presence of DSS. This indicated that the homodimer and the heterodimer were the potential receptor for CiIFN.
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Huarong
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Wenqun
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Binhua
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Huiling
authors.initials: H
authors.lastname: Mao
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Qunhao
authors.initials: Q
authors.lastname: Hou
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Yichuan
authors.initials: Y
authors.lastname: Mi
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Fan
authors.affiliation: Department of Bioscience, College of Life Science, Nanchang University, Nanchang 330031, China. Electronic address: hucy2008@163.com.
authors.forename: Chengyu
authors.initials: C
authors.lastname: Hu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D017471:Receptors, Interferon / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; D017471:Receptors, Interferon
keywords: Antiviral signaling pathway; CRFB1 and CRFB5; Cytokine receptor family B; Teleost; Type I interferon