Chen Si, He Hua, Liu Xiaolin
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
PLoS One. 2017 Apr 17;12(4):e0175777. doi: 10.1371/journal.pone.0175777. eCollection 2017.
In mammals, very long chain fatty acids (VLCFAs) perform pleiotropic roles in a wide range of biological processes, such as cell membrane formation, cell signal transduction, and endocrine regulation. Beef and milk are abundant of palmitic acid which can be further elongated into stearic acid for synthesizing VLCFAs. Elongase of very long chain fatty acid 6 (ELOVL6) is a rate-limiting enzyme for converting palmitic acid to stearic acid. Consequently, investigating the tissue expression patterns and transcriptional regulation of bovine ELOVL6 can provide new insights into improving the composition of beneficial fats in cattle and expanding the knowledge of transcriptional regulation mechanism among domestic animals. In the current study, we found that bovine ELOVL6 expressed ubiquitously. Dual-luciferase reporter assay identified that the core promoter region (-130/-41 bp) was located in the second CpG island. In addition, the deletion mutation of binding sites demonstrated that sterol regulatory element binding transcription factor 1 (SREBF1) and specific protein 1 (SP1) both were able to stimulate bovine ELOVL6 promoter activity independently, while resulting the similar effect. To confirm these findings, further RNA interference assays were executed in bovine mammary epithelial cells (BMECs). In summary, these data suggest that bovine ELOVL6 expressed ubiquitously and is activated by SREBF1 and SP1, via two binding sites present in the ELOVL6 promoter region between -130 bp to -41bp.
在哺乳动物中,超长链脂肪酸(VLCFA)在广泛的生物过程中发挥着多效性作用,如细胞膜形成、细胞信号转导和内分泌调节。牛肉和牛奶富含棕榈酸,棕榈酸可进一步延长为硬脂酸用于合成超长链脂肪酸。超长链脂肪酸延长酶6(ELOVL6)是将棕榈酸转化为硬脂酸的限速酶。因此,研究牛ELOVL6的组织表达模式和转录调控可为改善牛体内有益脂肪的组成以及拓展家畜转录调控机制的知识提供新的见解。在本研究中,我们发现牛ELOVL6在全身广泛表达。双荧光素酶报告基因检测确定核心启动子区域(-130/-41 bp)位于第二个CpG岛。此外,结合位点的缺失突变表明,固醇调节元件结合转录因子1(SREBF1)和特异性蛋白1(SP1)均能独立刺激牛ELOVL6启动子活性,且效果相似。为证实这些发现,我们在牛乳腺上皮细胞(BMEC)中进行了进一步的RNA干扰试验。总之,这些数据表明牛ELOVL6在全身广泛表达,并通过ELOVL6启动子区域-130 bp至-41 bp之间存在的两个结合位点被SREBF1和SP1激活。