Identification of Fatty Acid Desaturase 6 in Golden Pompano Trachinotus Ovatus (Linnaeus 1758) and Its Regulation by the PPAR alpha b Transcription Factor

文献类型： 外文期刊

第一作者： Zhu, Ke-Cheng

作者： Zhu, Ke-Cheng;Song, Ling;Guo, Hua-Yang;Guo, Liang;Zhang, Nan;Liu, Bao-Suo;Jiang, Shi-Gui;Zhang, Dian-Chang;Zhu, Ke-Cheng;Guo, Hua-Yang;Guo, Liang;Zhang, Nan;Liu, Bao-Suo;Jiang, Shi-Gui;Zhang, Dian-Chang;Zhu, Ke-Cheng;Guo, Liang;Zhang, Nan;Liu, Bao-Suo;Zhang, Dian-Chang;Jiang, Shi-Gui

作者机构：

关键词： Trachinotus ovatus; fatty acid desaturases; PPAR alpha b and Fads6; transcriptional activity; heterologous expression

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2019 年 20 卷 1 期

页码：

收录情况： SCI

摘要： Fatty acid desaturases are rate-limiting enzymes in long-chain polyunsaturated fatty acid biosynthesis. The transcription factor peroxisome proliferator-activated receptor alpha b (PPAR alpha b) regulates lipid metabolism in mammals, however, the mechanism whereby PPAR alpha b regulates fatty acid desaturases is largely unknown in fish. In this study, we report the full length cDNA sequence of Trachinotus ovatus fatty acid desaturase, which encodes a 380 amino acid polypeptide, possessing three characteristic histidine domains. Phylogenetic and gene exon/intron structure analyses showed typical phylogeny: the T. ovatus fatty acid desaturase contained a highly conserved exon/intron architecture. Moreover, functional characterization by heterologous expression in yeast indicated that T. ovatus desaturase was a fatty acid desaturase, with Delta 4/Delta 5/Delta 8 Fad activity. Promoter activity assays indicated that ToFads6 desaturase transcription was positively regulated by PPAR alpha b. Similarly, PPAR alpha b RNA interference decreased ToPPAR alpha b and ToFads6 expression at the mRNA and protein levels in a time-dependent manner. Mutation analyses showed that the M2 binding site of PPAR alpha b was functionally important for protein binding, and transcriptional activity of the ToFads6 promoter was significantly decreased after targeted mutation of M2. Electrophoretic mobile shift assays confirmed that PPAR alpha b interacted with the binding site of the ToFads6 promoter region, to regulate ToFads6 transcription. In summary, PPAR alpha b played a vital role in ToFads6 regulation and may promote the biosynthesis of long-chain polyunsaturated fatty acids by regulating ToFads6 expression.

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