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首页> 外文期刊>Journal of dairy science >Overexpression of SREBF chaperone (SCAP) enhances nuclear SREBP1 translocation to upregulate fatty acid synthase (FASN) gene expression in bovine mammary epithelial cells
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Overexpression of SREBF chaperone (SCAP) enhances nuclear SREBP1 translocation to upregulate fatty acid synthase (FASN) gene expression in bovine mammary epithelial cells

机译：SREBF伴侣（SCAP）的过表达增强核SREBP1易位，以上调牛乳腺上皮细胞中的脂肪酸合酶（FASN）基因表达

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摘要

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.
机译：脂肪酸合酶是反刍动物乳腺中合成乳脂的关键酶。在非反刍动物中，固醇调节元件结合蛋白1（SREBP1）是FASN基因表达的调节剂，而SREBF分子伴侣（SCAP）对于SREBP1的成熟和活性至关重要。但是，SCAP在反刍动物中FASN基因表达调控中的作用尚不清楚。这项研究的目的是通过在牛乳腺上皮细胞中过度表达SCAP来研究FASN的转录调控。克隆了牛SCAP表达载体，SREBP1表达载体和FASN的启动子。使用生物信息学分析预测FASN启动子的转录因子结合位点。在永生化的牛乳腺上皮细胞系MAC-T中用FASN启动子载体转染后，我们以pcDNA3.1载体为对照共表达SCAP SREBP1表达载体。使用荧光素酶测定，免疫荧光，蛋白质印迹，实时PCR和脂质滴染色研究了SCAP SREBP1过表达对FASN调节的影响。我们观察到，SCAP SREBP1的共过量表达显着增加了含有固醇反应元件结合位点的FASN启动子的活性。由于SCAP SREBP1的共过量表达，FASN mRNA的丰度和脂质滴的形成增加了。与单独的SREBP1的过表达相比，SCAP SREBP1的共过表达增强了核易位和核SREBP1蛋白的丰度。总体而言，与非反刍动物细胞一样，结果表明SCAP对于促进SREBP1的核易位和FASN基因转录的激活至关重要，从而导致牛乳腺上皮细胞中脂质滴的形成。

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  • 来源
    《Journal of dairy science》 |2018年第7期|6523-6531|共9页
  • 作者

    Han L. Q.; Gao T. Y.; Yang G. Y.; Loor J. J.;

  • 作者单位

    Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Henan, Peoples R China;

    Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Henan, Peoples R China;

    Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Henan, Peoples R China;

    Univ Illinois, Dept Anim Sci, 328 Mumford Hall, Urbana, IL 61801 USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    lactation; lipogenesis; milk fat;

    机译：泌乳;脂肪生成;乳脂;

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