Transcriptional regulation of porcine PABPN1 gene in adipogenesis.

Abstract

Objective: This study was designed to reveal the transcriptional regulatory mechanism and effects of poly(A)-binding protein nuclear 1 (PABPN1)on adipogenesis and the polymorphisms as well.

Methods: Transcription factors were identified with dual-luciferase reporter assay, overexpression, site-directed mutagenesis, real-time quantitative PCR (qPCR), electrophoretic mobility shift assay and Chromatin immunoprecipitation-qPCR. Preadipocyte differentiation was measured with gain- and loss-of-function, Oil Red O staining and extraction assays. Single nucleotide polymorphisms (SNPs) were identified with direct sequencing of PCR products in the promoter, and the effects of SNPs on PABPN1 expression were identified with dual-luciferase reporter assay.

Results: Both CCAAT/enhancer-binding protein (C/EBP) α and β can regulate the expression of PABPN1 by directly binding to the promoter. PABPN1 promotes the preadipocyte differentiation in pigs. A total of three SNPs were identified, and the haplotype mutation, Haplotype GCC, significantly increases the promoter activity of PABPN1.

Conclusions: PABPN1 promotes the preadipocyte differentiation as a downstream gene of C/EBP α and β. Haplotype GCC has the potential as a molecular marker for selecting fat trait in pigs.