Characteristics of the interferon regulatory factor IRF5 and its stimulation expression by Elizabethkingia miricola Infection in black-spotted frog (Pelophylax nigromaculatus)

Abstract

Interferon regulatory factors (IRFs) have been confirmed as vital transcription factors for the immune system, which play a certain role in anti-infection defense, immune regulation, hematopoietic cell development, and maturation of the immune system. In this study, the cDNA of Interferon regulatory factor 5 (IRF5) was cloned from black-spotted frog, Pelophylax nigromaculatus, and termed PnIRF5. The results indicated, the full-length cDNA of PnIRF5 was 2090 bp, and a putative protein of 504 amino acids was encoded. The results of protein domain prediction suggested that PnIRF5 exhibited a DNA-binding domain (DBD), a middle region (MR), an IRF association domain (IAD), a virus activated domain (VAD), as well as two nuclear localization signals (NLSs). Phylogenetic analysis clustered PnIRF5 into the amphibian IRF5 subgroup in vertebrate IRF5 group. The results of qPCR show that PnIRF5 mRNA was expressed in all examined tissues, with higher levels were identified in the kidney, intestine, and lung. The gene expression of PnIRF5 was analyzed in kidney, spleen, and liver of black-spotted frog after challenged with Elizabethkingia miricola, and the maximum expression level of PnIRF5 in liver, spleen, and kidney were 1.80, 2.33 and 2.88 times that at 0 days post challenged, respectively. As indicated by the results, the PnIRF5 protein may take on critical significance in protecting black-spotted frog from E. miricola infection, and they may play a certain role in the immune response of black-spotted frog against bacteria.