The Investigation Of The Transcription Regulation And Signaling Pathway Of Porcine S100A12Gene

Haemophilus parasuis(HPS) cause HPS disease after infecting pigs. Its high morbidity and mortality made it become one of the common bacteria in the world wide. Up to now, we know little about the molecular mechanism and host response when pigs infected with HPS. Transcriptome sequencing data of pig spleen after HPS infection showed S100A8, S100A9and S100A12gene were significantly upregulated, the same as using LPS and poly I:C stimulation. As these three genes shared the same expression patterns, we focus on pig S100A12gene to study the transcriptional regulation and signal pathway in order to reveal the functions of these three genes. The main results are as follows:1. We investigated the transcriptional regulation of pig S100A12by Dual-Luciferase Reporter assay system, the results showed that pig S100A12was negatively regulated by Sox-5.2. RT-PCR was used to investigate the expression patterns of pig S100A8, S100A9, S100A12and RAGE gene, results showed pig S100A8, S100A9, S100A12were highly expressed in spleen and lung, and they shared the same expression patterns. Pig RAGE gene was strictly expressed in pig lung tissue.3. In PK-15cell line, S100A12engaged with RAGE can not activate NF-κB, but S100A12can activate NF-κB through TLR4receptor.4. In C2C12cell line, S100A12engaged with RAGE can activate NF-κB, and there existed a dose-dependent effect. S100A8/A9engaged with RAGE decreased the activity of NF-κB.5. We found RAGE can interact with NCK1, GrB2, RHOA in the cell membrane by BIFC method, and NCK1was mainly expressed in the cytoplasm, GrB2was mainly expressed in the nucleus, RHOA was expressed in the whole cell.