Molecular characterization of interferon regulatory factor 2 (IRF-2) homolog in pearl oyster Pinctada fucata

• Published in: Fish & shellfish immunology Vol. 34; no. 5; pp. 1279 - 1286
• Main Authors: Huang, Xian-De, Liu, Wen-Guang, Wang, Qi, Zhao, Mi, Wu, Shan-Zeng, Guan, Yun-Yan, Shi, Yu, He, Mao-Xian
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.05.2013
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA, Complementary - genetics; DNA, Complementary - metabolism; Fluorescent Antibody Technique; Gene cloning; Gene Expression Regulation; HEK293 Cells; Humans; Immune response; Immunity, Innate; Interferon Regulatory Factor-2 - chemistry; Interferon Regulatory Factor-2 - genetics; Interferon Regulatory Factor-2 - immunology; Lipopolysaccharides - pharmacology; Molecular Sequence Data; NF-kappa B - metabolism; Organ Specificity; Pearl oyster; PfIRF-2; Phylogeny; Pinctada - chemistry; Pinctada - genetics; Pinctada - immunology; Poly I-C - pharmacology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Alignment; Pearl oyster; Gene cloning; Immune response; PfIRF-2
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2013.02.003

Interferon regulatory factors (IRFs) control many facets of the innate and adaptive immune responses, regulate the development of the immune system itself and involve in reproduction and morphogenesis. In the present study, the IRF-2 homology gene, PfIRF-2 from pearl oyster Pinctada fucata was cloned and its genomic structure and promoter were analyzed. PfIRF-2 encodes a putative protein of 350 amino acids, and contains a highly conserved N-terminal DNA-binding domain and a variable C-terminal regulatory domain. Comparison and phylogenetic analysis revealed that PfIRF-2 shared a relatively higher identity with other mollusk but relatively lower identity with vertebrate IRF-2, and was clustered with IRF-1 subfamily composed of IRF-2 and IRF-1. Furthermore, gene expression analysis revealed that PfIRF-2 involved in the immune response to LPS and poly(I:C) stimulation. Immunofluorescence assay showed that the expressed PfIRF-2 was translocated into the nucleus and dual-luciferase reporter assays indicated that PfIRF-2 could involved and activate interferon signaling or NF-κB signal pathway in HEK293 cells. The study of PfIRF-2 may help better understand the innate immune in mollusk. ► Interferon regulatory factors 2 gene was isolated and characterized from Pinctada fucata. ► PfIRF-2 clustered with IRF-1 subfamily composed of IRF-2 and IRF-1. ► PfIRF-2 involved in the immune response to LPS and poly(I:C) stimulation. ► The expressed PfIRF-2 was translocated into the nucleus. ► PfIRF-2 could active target genes containing the IRSE or NF-κB binding site in vitro.