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大黄花鱼(Larimichthys crocea)中假定的 Elovl5 和 Elovl4 延长酶的功能特征和差异营养调控。

Functional characterization and differential nutritional regulation of putative Elovl5 and Elovl4 elongases in large yellow croaker (Larimichthys crocea).

机构信息

Key Laboratory of Aquaculture Nutrition and Feed (Ministry of Agriculture) and Key Laboratory of Mariculture (Ministry of Education), Ocean University of China, Qingdao, 266003, People's Republic of China.

Institute of Aquaculture, School of Natural Sciences, University of Stirling, Stirling, FK9 4LA, Scotland, UK.

出版信息

Sci Rep. 2017 May 23;7(1):2303. doi: 10.1038/s41598-017-02646-8.

Abstract

In the present study, two elongases, Elovl4 and Elovl5, were functionally characterized and their transcriptional regulation in response to n-3 LC-PUFA administration were investigated in vivo and in vitro. We previously described the molecular characterization of croaker elovl5. Here, we report the full-length cDNA sequence of croaker elovl4, which contained 1794 bp (excluding the polyA tail), including 909 bp of coding region that encoded a polypeptide of 302 amino acids possessing all the characteristic features of Elovl proteins. Functional studies showed that croaker Elovl5, displayed high elongation activity towards C and C PUFA, with only low activity towards C PUFA. In contrast, croaker Elovl4 could effectively convert both C and C PUFA to longer polyenoic products up to C. n-3 LC-PUFA suppressed transcription of the two elongase genes, as well as srebp-1 and lxrα, major regulators of hepatic lipid metabolism. The results of dual-luciferase reporter assays and in vitro studies both indicated that the transcriptions of elovl5 and elovl4 elongases could be regulated by Lxrα. Moreover, Lxrα could mediate the transcription of elovl4 directly or indirectly through regulating the transcription of srebp-1. The above findings contribute further insight and understanding of the mechanisms regulating LC-PUFA biosynthesis in marine fish species.

摘要

在本研究中,我们对两种延长酶(Elovl4 和 Elovl5)进行了功能表征,并研究了它们在体内和体外对 n-3LC-PUFA 给药的转录调控。我们之前描述了黄鱼 Elovl5 的分子特征。在这里,我们报告了黄鱼 Elovl4 的全长 cDNA 序列,其包含 1794bp(不包括 polyA 尾巴),包括 909bp 的编码区,编码一个由 302 个氨基酸组成的多肽,具有 Elovl 蛋白的所有特征。功能研究表明,黄鱼 Elovl5 对 C 和 C PUFA 具有高延长活性,而对 C PUFA 的活性仅低。相比之下,黄鱼 Elovl4 可以有效地将 C 和 C PUFA 转化为更长的多烯产物,直到 C。n-3LC-PUFA 抑制了两种延长酶基因以及肝脏脂质代谢的主要调节剂 Srebp-1 和 Lxrα 的转录。双荧光素酶报告基因检测和体外研究的结果均表明,Lxrα 可以调节 Elovl5 和 Elovl4 延长酶的转录。此外,Lxrα 可以通过调节 Srebp-1 的转录来直接或间接调节 Elovl4 的转录。上述发现为海洋鱼类 LC-PUFA 生物合成的调控机制提供了进一步的深入了解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a70c/5442133/4007a7439cd6/41598_2017_2646_Fig1_HTML.jpg

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