Abstract

The proinflammatory cytokine, interleukin-6 (IL-6), plays a critical role in many chronic inflammatory diseases, particularly inflammatory bowel disease. To investigate the regulation of IL-6 gene expression at the molecular level, genomic DNA sequencing of Jinghai yellow chickens (Gallus gallus) was performed to detect single-nucleotide polymorphisms (SNPs) in the region −2200 base pairs (bp) upstream to 500 bp downstream of IL-6. Transcription factor binding sites and CpG islands in the IL-6 promoter region were predicted using bioinformatics software. Twenty-eight SNP sites were identified in IL-6. Four of these 28 SNPs, three [−357 (G > A), −447 (C > G), and −663 (A > G)] in the 5′ regulatory region and one in the 3′ non-coding region [3177 (C > T)] are not labelled in GenBank. Bioinformatics analysis revealed 11 SNPs within the promoter region that altered putative transcription factor binding sites. Furthermore, the C-939G mutation in the promoter region may change the number of CpG islands, and SNPs in the 5′ regulatory region may influence IL-6 gene expression by altering transcription factor binding or CpG methylation status. Genetic diversity analysis revealed that the newly discovered A-663G site significantly deviated from Hardy-Weinberg equilibrium. These results provide a basis for further exploration of the promoter function of the IL-6 gene and the relationships of these SNPs to intestinal inflammation resistance in chickens.

Highlights

  • Interleukin-6 (IL-6), known as interferon β-2, stem cell-stimulating factor, or B cell differentiation factor, is a multifunctional cytokine that plays an important role in regulating the immune response, acute-phase response, and haematopoiesis in chickens [1]

  • A total of 28 single-nucleotide polymorphisms (SNPs) sites were detected (Table 2), among which 19 SNPs are located in the 50 regulatory region, and three were newly discovered; the chromosomal positions of these three SNPs are 30948722, 30948938, and 30949028

  • To determine the effect of these SNPs on changes of the predicted CpG island in the promoter region of IL-6, MethPrimer software was used to predict the presence of CpG islands in the 2200-bp sequence upstream of the transcription start site of the IL-6 promoter region before and after SNP site mutations

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Summary

Introduction

Interleukin-6 (IL-6), known as interferon β-2, stem cell-stimulating factor, or B cell differentiation factor, is a multifunctional cytokine that plays an important role in regulating the immune response, acute-phase response, and haematopoiesis in chickens [1]. Amrani et al [2]. First discovered that chicken IL-6 influences hepatocyte-stimulating factor (HSF) in the acute-phase response. Schneider et al [3] successfully cloned the complete reading frame cDNA for chicken IL-6. Horiuchi et al [4] reported that the chicken leukaemia inhibitory factor belongs to the IL-6 family. By injecting DNA plasmids carrying the very virulent infectious bursal disease virus (vvIBDV) SH95 poly-protein (VP2-4-3) gene and DNA plasmids carrying chicken IL-6 (ChIL-6), Sun et al [5] determined that injection with ChIL-6 plasmids resulted in a significantly increased protective effect in chickens after infection with highly virulent strains. IL-6 plays a vital role in the chronic inflammation and

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