A 914-bp promoter is sufficient to reproduce the endogenous prolyl oligopeptidase gene localization in the mouse placenta if not subject to position effect

Abstract

Prolyl oligopeptidase (POP) is a widely distributed multifunctional protein which has an endopeptidase activity to cleave a -Pro-X- peptide bond. In spite of numerous studies about POP, the mechanism by which its transcription is controlled has not been well investigated. Here we generated transgenic mice bearing a transgene which contained a 914-bp POP gene promoter linked to the enhanced green fluorescent protein (EGFP) gene to assess the in vivo promoter activity. We established six transgenic lines with different copy numbers, but no EGFP signal was observed in four lines due to a high level of DNA methylation, which suggested that the transgene was subject to position effect. However, in the other two lines, we detected the EGFP expression in many tissues, and its placental localization showed a similar change to POP. A strong EGFP signal was observed in the junctional and labyrinthine zones of E10.5-E12.5 placentas and in the junctional zone and the maternal decidua after that. This placental gene activation might be attributed to AP-2 gamma because we detected its binding to the POP promoter. In contrast, we did not obtain any evidence that EGFP was expressed in a similar pattern compared with POP in the ovary. The current data demonstrated that the 914-bp promoter had sufficient activity to reproduce the POP localization in the placenta if it was not subject to position effect and suggest that the regulatory mechanism of the POP gene expression differs between tissues.(C) 2013 Elsevier B.V. All rights reserved

A molecular mechanism of mouse placental spongiotrophoblast differentiation regulated by prolyl oligopeptidase

“…POP could control Ascl2 by changing the phosphorylation status of proteins in these pathways, as it has been reported to regulate protein phosphorylation (Williams et al 1999; Duan et al 2014). It is interesting that Ascl2 is activated by a placenta-specific transcription factor, AP-2γ (Sharma et al 2016), and POP is also suggested to be controlled by AP-2γ (Matsubara et al 2013). SpT differentiation may be regulated by an AP-2γ–POP–Ascl2 pathway.…”

A Long Noncoding RNA, lncRNA-Amhr2, Plays a Role in Amhr2 Gene Activation in Mouse Ovarian Granulosa Cells

“…6.1k-POPpr-Luc: A 914-bp mouse prolyl oligopeptidase (POP) gene promoter was subcloned into a pEGFP-1 vector (Clontech) (47,48). The plasmid was digested with BamHI and EcoRI, and the POP promoter was purified, blunted, and inserted into 6.1k-Luc which was digested with EcoRV and Tth111I and blunted.…”

“…Then, we replaced most of the lncRNA-Amhr2 sequence with torazame ER cDNA which was similar in length (6.1k-toER-Luc) (46), and the lncRNA-Amhr2 promoter sequence with the POP promoter (6.1k-POPpr-Luc). The replacement with torazame ER would result in transcription of a sequence unrelated to lncRNA-Amhr2, and the POP promoter was known to show strong promoter activity in various types of cells and expected to drive lncRNA-Amhr2 transcription at a higher level than other constructs (47,48,51). As controls, we also prepared the constructs that contained the 0.5-kb Amhr2 promoter (0.5k-Luc) and lacked a lncRNA-Amhr2 promoter sequence (6.1k-∆lncPr-Luc).…”