title: Molecular characterization of the porcine S100A6 gene and analysis of its expression in pigs infected with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV).
abstract: Our previous microarray study revealed that S100A6 was significantly upregulated in porcine alveolar macrophages (PAMs) infected with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). In the present study, we cloned both cDNA and genomic DNA sequences of the gene. Transient transfection indicated that the porcine S100A6 protein was located in the nucleus and cytoplasm. Reverse transcription polymerase chain reaction (RT-PCR) revealed that the porcine S100A6 gene was highly expressed in the kidney and subcutaneous fat. Polyinosinic-polycytidylic acid [poly (I:C)] induced porcine S100A6 gene expression in PK-15 cells. Quantitative real-time PCR (Q-PCR) analysis further showed that the porcine S100A6 gene was upregulated in different cells and tissues of Tongcheng pigs infected with HP-PRRSV. Chromosome walking obtained the porcine S100A6 promoter region and then luciferase reporter assays confirmed its regulatory activities. We observed a putative NF-κB binding site in the core promoter region, which may explain the upregulation of porcine S100A6 in response to PRRSV. Transfection of NF-κB (p65 subunit) intensely induced the promoter activity of the porcine S100A6 gene, while an NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), inhibited this activity. Furthermore, compared to its wild type, the promoter activity was significantly reduced when it contained a mutant NF-κB binding site. All these results provide a solid foundation to further investigate how S100A6 is involved in PRRSV infection.
authors.affiliation: Key Lab of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhou
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Michal
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Jinhua
authors.initials: J
authors.lastname: Zhao
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Jiang
authors.forename: Bang
authors.initials: B
authors.lastname: Liu
mesh_terms: D000818:Animals; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular; D007668:Kidney / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D019318:Porcine Reproductive and Respiratory Syndrome / Q000235:genetics*; D019316:Porcine respiratory and reproductive syndrome virus; D011401:Promoter Regions, Genetic; D009418:S100 Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA; D050151:Subcutaneous Fat / Q000378:metabolism; D034421:Sus scrofa / Q000235:genetics*; D013552:Swine
chemical_list: D018797:Cell Cycle Proteins; D016328:NF-kappa B; D009418:S100 Proteins