The miR-15/16 Cluster Is Involved in the Regulation of Vertebrate LC-PUFA Biosynthesis by Targeting ppar gamma as Demonstrated in Rabbitfish Siganus canaliculatus
The miR-15/16 Cluster Is Involved in the Regulation of Vertebrate LC-PUFA Biosynthesis by Targeting ppar gamma as Demonstrated in Rabbitfish Siganus canaliculatus
复制标题
miR-15/16 簇通过靶向 ppar gamma 参与脊椎动物 LC-PUFA 生物合成的调节，如兔鱼 Siganus canaliculatus 中所示
DOI：
10.1007/s10126-020-09969-0
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发表时间：
2020
影响因子：
3
通讯作者：
Li Yuanyou
中科院分区：
文献类型：
--
作者：
Sun Junjun;Chen Cuiying;You Cuihong;Liu Yang;Ma Hongyu;Monroig Oscar;Tocher Douglas R.;Wang Shuqi;Li Yuanyou
转录后调控机制在长链（≥ C20）多不饱和脂肪酸（LC-PUFA）生物合成的调控中起重要作用。在这里，我们解决了一个潜在的重要作用的miR-15/16簇在调节LC-PUFA的生物合成在rabbitfishSiganus canaliculatus。在兔鱼中，miR-15和miR-16都对影响LC-PUFA生物合成的脂肪酸高度响应，并且在一系列兔鱼组织中显示出相似的表达模式。在兔鱼LC-PUFA生物合成的抑制剂过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor gamma，pparγ）的3′UTR中预测了miR-15和miR-16共同的潜在结合位点，荧光素酶报告基因分析表明ppar γ是miR-15/16簇的潜在靶点。在体外实验中，miR-15和miR-16单独或联合过表达可抑制Pparγ的mRNA和蛋白水平，并增加LC-PUFA合成关键酶Δ6Δ5 fads 2、Δ4 fads 2和delov 15的mRNA水平。在SCHL中，miR-15和miR-16共同过表达对ppar γ的抑制作用比单独过表达更明显。敲低miR-15/16簇得到相反的结果，并且在敲低ppar γ后观察到LC-PUFA生物合成酶的mRNA水平增加。miR-15/16簇过表达显著增加SCHL中22：6 n-3、20：4 n-6和总LC-PUFA的含量，其中18：4 n-3/18：3 n-3和22：6 n-3/22：5 n-3比值更高。这些结果表明，miR-15和miR-16作为一个miRNA簇，共同作用于ppar γ，促进了兔鱼LC-PUFA的合成。这是首次报道miR-15/16簇参与脊椎动物LC-PUFA生物合成。
Post-transcriptional regulatory mechanisms play important roles in the regulation of long-chain (≥ C20) polyunsaturated fatty acid (LC-PUFA) biosynthesis. Here, we address a potentially important role of the miR-15/16 cluster in the regulation of LC-PUFA biosynthesis in rabbitfishSiganus canaliculatus. In rabbitfish, miR-15 and miR-16 were both highly responsive to fatty acids affecting LC-PUFA biosynthesis and displayed a similar expression pattern in a range of rabbitfish tissues. A common potential binding site for miR-15 and miR-16 was predicted in the 3′UTR of peroxisome proliferator-activated receptor gamma (pparγ), an inhibitor of LC-PUFA biosynthesis in rabbitfish, and luciferase reporter assays revealed thatpparγwas a potential target of miR-15/16 cluster. In vitro individual or co-overexpression of miR-15 and miR-16 in rabbitfish hepatocyte line (SCHL) inhibited both mRNA and protein levels of Pparγ, and increased the mRNA levels ofΔ6Δ5 fads2,Δ4 fads2, andelovl5, key enzymes of LC-PUFA biosynthesis. Inhibition ofpparγwas more pronounced with co-overexpression of miR-15 and miR-16 than with individual overexpression in SCHL. Knockdown of miR-15/16 cluster gave opposite results, and increased mRNA levels of LC-PUFA biosynthesis enzymes were observed after knockdown ofpparγ. Furthermore, miR-15/16 cluster overexpression significantly increased the contents of 22:6n-3, 20:4n-6 and total LC-PUFA in SCHL with higher 18:4n-3/18:3n-3 and 22:6n-3/22:5n-3 ratio. These suggested that miR-15 and miR-16 as a miRNA cluster together enhanced LC-PUFA biosynthesis by targetingpparγin rabbitfish. This is the first report of the participation of miR-15/16 cluster in LC-PUFA biosynthesis in vertebrates.