Molecular insight, expression profile and subcellular localization of two STAT family members, STAT1a and STAT2, from Japanese eel, Anguilla japonica

Abstract

Signal transducer and activator of transcription 1 (STAT1) and STAT2 are critical components of type I and type II IFNs signaling. To date, seven STAT family proteins have been identified from mammals. However, the information on STAT genes in teleost fish is still limited. In the present study, two STAT family genes (STAT1a and STAT2) were identified from Japanese eel, Anguilla japonica and designated as AjSTAT1a and AjSTAT2. The open reading frames of AjSTAT1a and AjSTAT2 are 2244 bp and 2421 bp, encoding for polypeptides of 747 aa and 806 aa, respectively. Both AjSTAT1a and AjSTAT2 contain the conserved domains of STAT proteins. Phylogenetic analysis was performed based on the STATs protein sequences, and showed that AjSTAT1a and AjSTAT2 shared the closest relationship with Oncorhynchus mykiss. Quantitative real-time PCR analysis revealed that AjSTAT1a and AjSTAT2 were expressed in most examined tissues, with the highest expression both in blood. Significantly up-regulated transcripts of AjSTAT1a and AjSTAT2 were detected in response to poly I:C stimulation, and Edwardsiella tarda induced increase in the expression of AjSTAT1a and AjSTAT2 genes. Subcellular localization analysis showed that in both IFNγ-stimulated and unstimulated EPC cells AjSTAT1a and AjSTAT2 were mainly distributed in the cytoplasm, but few AjSTAT1a was distributed in the nucleus. All these results suggested that AjSTAT1a and AjSTAT2 may be critical for regulating the host innate immune defense against pathogens invasion.

Keywords: Anguilla japonica; Expression; STAT1a; STAT2; Subcelluar localization.