One SNP in the 3′-UTR of HMGB1 gene affects the binding of target bta-miR-223 and is involved in mastitis in dairy cattle

Abstract

High-mobility group box protein 1 (HMGB1) gene has a universal sentinel function for nucleic acid-mediated innate immune responses and acts as a pathogenic mediator in the inflammatory disease. In an effort to identify the functional single-nucleotide polymorphism (SNP) in the 3'-untranslated region (UTR) of the bovine HMGB1 gene that affects the binding to its target microRNA, first, the expression of HMGB1 mRNA in different genotypes and its candidate bta-miR-223 was investigated. Quantitative real-time polymerase chain reaction results showed that the relative expression of HMGB1 mRNA in cows with the genotype GG is significantly higher than those in cows with the genotype AA (P < 0.05). The expression of bta-miR-223 was significantly upregulated by 1.95-fold (P < 0.05) in the bovine mastitis-infected mammary gland tissues compared with that in the healthy tissues. Subsequently, luciferase assay indicated that the HMGB1 expression was directly targeted by bta-miR-223 in human embryo kidney 293 T (HEK 293T) cells. One novel SNP (g. +2776 A > G) in the HMGB1 3'-UTR, altering the binding of HMGB1 and bta-miR-223, was found to be associated with somatic count scores in cows. Taken together, the g. +2776 A > G-GG was an advantageous genotype which can be used as a candidate functional marker for mastitis resistance breeding program.

Genome-Wide Specific Selection in Three Domestic Sheep Breeds

“…CIB2 is associated with influencing interleukin levels in African Americans [46]. HMGB1 is involved in mastitis in dairy cattle [47]. TRIOBP and TMC2 can cause recessive hearing loss in human [48, 49].…”

The Role of microRNAs in Bovine Infection and Immunity

“…One example where miRNA regulation has been validated is bovine High Mobility Group Box 1 (HMGB1), a nuclear protein, which transcriptionally regulates inflammation (48). Bovine HMGB1 has been shown to be targeted by bta-miR-223, a miRNA that is up-regulated during S. aureus infection (48).…”

“…One example where miRNA regulation has been validated is bovine High Mobility Group Box 1 (HMGB1), a nuclear protein, which transcriptionally regulates inflammation (48). Bovine HMGB1 has been shown to be targeted by bta-miR-223, a miRNA that is up-regulated during S. aureus infection (48). Similarly, using reporter assays, bta-miR-124 has been shown to regulate expression of Monocyte Chemotactic Protein 1 (MCP1) and Polypyrimidine Tract Binding Protein 1 (PTBP1) in bovine fibroblasts (53).…”

Genetic mutations potentially cause two novel NCF1 splice variants up-regulated in the mammary gland, blood and neutrophil of cows infected by Escherichia coli

“…Five DNA pools were used to amplify the fragments with the above primers and then PCR products were sent to the Invitrogen Company (Invitrogen, Beijing, China) for direct sequencing. Genotyping of SNP was determined by PCR-RFLP as described by Li et al (2012). Briefly, the restriction endonucleases (Nla III, Bsh1285I and FnuD II, Fermentas, Canada) were selected to digest PCR products and the digested fragments were separated by polyacrylamide gel electrophoresis (PAGE) using 8% polyacrylamide gels.…”