A RING finger protein 114 (RNF114) homolog from Chinese sturgeon (Acipenser sinensis) possesses immune-regulation properties via modulating RIG-I signaling pathway-mediated interferon expression

• Published in: Fish & shellfish immunology 2014-12, Vol.41 (2), p.507-516
• Main Authors: Liao, Zhiyong, Chen, Xiaojun, Nie, Dongsong, Wang, Jiajia, Wu, Mingjiang
• Format: Article
• Language: English
• Subjects: Acipenser; Acipenser sinensis; Analysis of Variance; Animals; cDNA cloning; Chinese sturgeon; Cloning, Molecular; Computational Biology; DNA Primers - genetics; Fishes - genetics; Fishes - immunology; Gene Knockdown Techniques; Gene Library; HEK293 Cells; Humans; Immune response; Immunoprecipitation; Interferons - metabolism; NF-kappa B - metabolism; Open Reading Frames - genetics; Phylogeny; Poly I-C; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RIG-I; RING Finger Domains - genetics; RING Finger Domains - immunology; RNF114; Signal Transduction - immunology; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - immunology; Ubiquitin-Protein Ligases - metabolism
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2014.09.030

Ubiquitin ligases play important roles in immune regulation. The human RNF114 (RING finger protein 114), an ubiquitin ligase, was recently reported to be involved in immune response to double-stranded RNA in disease pathogenesis. Here, we identified a RNF114 homolog in Chinese sturgeon (Acipenser sinensis) and investigated its potential role in immune response. The full-length cDNA of Chinese sturgeon RNF114 (csRNF114) contains an open reading frame (ORF) of 681 nucleotides coding a protein of 227 amino acids. csRNF114 shares the highest identity of 76% at amino acid level to other RNF114 homologs, clustering with bony fish RNF114s based on phylogenetic analysis. The main structural features of csRNF114, including a C3HC4 (Cys3-His-Cys4) RING domain, a C2HC (Cys2-His-Cys)-type zinc finger motif, a C2H2 (Cys2-His2)-type zinc finger motif, and a UIM (ubiquitin-interacting motif), take csRNF114 as an ubiquitin ligase. csRNF114 mRNA was widely expressed in various tissues and significantly up-regulated in poly(I:C)-treated Chinese sturgeon. Over-expression of csRNF114 in HEK293T cells significantly promoted both basal and poly(I:C)-induced activation of interferon regulatory transcription factor 3 (IRF3) and nuclear factor-κB (NF-κB) downstream retinoic acid inducible gene I (RIG-I) signaling pathway and expression of target genes type I interferon (IFN), which was nearly abolished by knockdown of RIG-I with specific human siRNA and by mutation of the C3HC4 RING domain (C28A/C31A) in csRNF114 as well. Furthermore, csRNF114 associated with ubiquitinated proteins in HEK293T cells, for which the C3HC4 RING domain was essential. These data suggested that an ubiquitin ligase RNF114 homolog with a potential role in antiviral response possibly through modulating RIG-I signaling pathway was cloned from Chinese sturgeon, which might contribute to our understanding of the immune biology of Chinese sturgeon. •A RNF114 homolog was cloned and identified in Chinese sturgeon.•csRNF114 shared distinct structural features of E3 and displayed E3 ligase activity.•Poly(I:C) stimulation induced csRNF114 expression in Chinese Sturgeon.•csRNF114 regulated interferon secretion in HEK293T cell via modulating RIG-I pathway.•The RING domain is essential to csRNF114 function in HEK293T cell.