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The hormone gastrin physiologically regulates gastric acid secretion and also contributes to maintaining gastric epithelial architecture by regulating expression of genes such as  plasminogen activator inhibitor 2 (PAI-2)  and  regenerating protein 1(Reg1) . Here we examine the role of proteasome subunit PSMB1 in the transcriptional regulation of  PAI-2  and  Reg1  by gastrin, and its subcellular distribution during gastrin stimulation. We used the gastric cancer cell line AGS, permanently transfected with the CCK2 receptor (AGS-GR) to study gastrin stimulated expression of  PAI-2  and  Reg1  reporter constructs when PSMB1 was knocked down by siRNA. Binding of PSMB1 to the  PAI-2  and  Reg1  promoters was assessed by chromatin immunoprecipitation (ChIP) assay. Subcellular distribution of PSMB1 was determined by immunocytochemistry and Western Blot. Gastrin robustly increased expression of  PAI-2  and  Reg1  in AGS-GR cells, but when PSMB1 was knocked down the responses were dramatically reduced. In ChIP assays, following immunoprecipitation of chromatin with a PSMB1 antibody there was a substantial enrichment of DNA from the gastrin responsive regions of the  PAI-2  and  Reg1  promoters compared with chromatin precipitated with control IgG. In AGS-GR cells stimulated with gastrin there was a significant increase in the ratio of nuclear:cytoplasmic PSMB1 over the same timescale as recruitment of PSMB1 to the  PAI-2  and  Reg1  promoters seen in ChIP assays. We conclude that PSMB1 is part of the transcriptional machinery required for gastrin stimulated expression of  PAI-2  and  Reg1 , and that its change in subcellular distribution in response to gastrin is consistent with this role.

The Role of Proteasome Beta Subunits in Gastrin-Mediated Transcription of Plasminogen Activator Inhibitor-2 and Regenerating Protein1