Characterization of the NLRC5 promoter in chicken: SNPs, regulatory elements and CpG islands

Abstract

NLRC5 plays an important role in the innate immunity and cellular immunity in many species, but the regulatory mechanism of NLRC5 expression in chickens remains unclear. In this study, a series of deletion fragments of the NLRC5 promoter region were constructed and dual-luciferase assay was performed. Then, we detected the SNP in the core region and its function. Important transcriptional regulatory elements were predicted and identified. Methylation of CpG islands was measured. The results revealed that the two core regions of -4372 to -3756 and -2925 to -2265 in the NLRC5 promoter were essential for NLRC5 mRNA expression in which a SNP (A/G), located at -2470, was found to have an effect on the transcriptional activity. Also, the STAT1 element in the second core region of the NLRC5 promoter was identified to bind with the STAT1 transcription factor, which was necessary for the transcriptional activity. In addition, many other elements in the NLRC5 promoter, including YY1 and CEBP, may contribute significantly to the expression activity of NLRC5. Moreover, two CpG islands were searched. Part of one was located in the first core region, which suggests that epigenetic modification may regulate the activity of the first promoter region, and the other was mostly in an unmethylated state. Collectively, these results suggest the complex regulation of NLRC5 expression includes SNPs, transcription factors and methylation.