Journal
MARINE BIOTECHNOLOGY
Volume 22, Issue 4, Pages 475-487Publisher
SPRINGER
DOI: 10.1007/s10126-020-09969-0
Keywords
miR-15/16 cluster; ppar gamma; Delta 6 Delta 5 fads2; Delta 4 fads2; LC-PUFA biosynthesis; Rabbitfish Siganus canaliculatus
Funding
- National Key R&D Program of China [2018YFD0900400]
- National Natural Science Foundation of China [31873040, 31702357]
- Natural Science Foundation of Guangdong Province [2020A1515011252, 2018A030313910]
- China Agriculture Research System [CARS-47]
- Innovation and Strong School Projects in Guangdong Province [2016KTSCX037]
- STU Scientific Research Foundation for Talents [NTF19019]
- Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding [2017B030314058]
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Post-transcriptional regulatory mechanisms play important roles in the regulation of long-chain (>= C-20) 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 rabbitfish Siganus 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 (ppar gamma), an inhibitor of LC-PUFA biosynthesis in rabbitfish, and luciferase reporter assays revealed that ppar gamma was 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 Ppar gamma, and increased the mRNA levels of Delta 6 Delta 5 fads2, Delta 4 fads2, and elovl5, key enzymes of LC-PUFA biosynthesis. Inhibition of ppar gamma was 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 of ppar gamma. 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 targeting ppar gamma in rabbitfish. This is the first report of the participation of miR-15/16 cluster in LC-PUFA biosynthesis in vertebrates.
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