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  • Title: Overexpression of SREBF chaperone (SCAP) enhances nuclear SREBP1 translocation to upregulate fatty acid synthase (FASN) gene expression in bovine mammary epithelial cells.
    Author: Han LQ, Gao TY, Yang GY, Loor JJ.
    Journal: J Dairy Sci; 2018 Jul; 101(7):6523-6531. PubMed ID: 29680640.
    Abstract:
    Fatty acid synthase is a key enzyme for the synthesis of milk fat in the ruminant mammary gland. In nonruminants, sterol regulatory element binding protein 1 (SREBP1) is a regulator of FASN gene expression, and SREBF chaperone (SCAP) is essential for SREBP1 maturation and activity. However, the role of SCAP on the regulation of FASN gene expression in ruminants is unknown. The objective of this study was to investigate the transcriptional regulation of FASN by overexpressing SCAP in bovine mammary epithelial cells. A bovine SCAP expression vector, SREBP1 expression vector, and the promoter of FASN were cloned. The transcription factor binding sites of FASN promoter were predicted using bioinformatics analysis. After transfection with FASN promoter vectors in the immortalized bovine mammary epithelial cell line MAC-T, we co-overexpressed the SCAP + SREBP1 expression vector with pcDNA3.1 vector as control. The effect of SCAP + SREBP1 overexpression on the regulation of FASN was investigated using luciferase assay, immunofluorescence, Western blot, real-time PCR, and lipid droplet staining. We observed that co-overexpression of SCAP + SREBP1 significantly increased activity of the FASN promoter containing a sterol response element binding site. The FASN mRNA abundance and lipid droplet formation increased due to co-overexpression of SCAP + SREBP1. Compared with overexpression of SREBP1 alone, co-overexpression of SCAP + SREBP1 enhanced the nuclear translocation and nuclear SREBP1 protein abundance. Overall, as in nonruminants cells, results indicate that SCAP is essential for promoting nuclear translocation of SREBP1 and activation of FASN gene transcription, leading to lipid droplet formation in bovine mammary epithelial cells.
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