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The miR-15/16 Cluster Is Involved in the Regulation of Vertebrate LC-PUFA Biosynthesis by Targeting pparγ as Demonostrated in Rabbitfish Siganus canaliculatus

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Abstract

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 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 (pparγ), an inhibitor of LC-PUFA biosynthesis in rabbitfish, and luciferase reporter assays revealed that pparγ 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 Pparγ, and increased the mRNA levels of Δ6Δ5 fads2, Δ4 fads2, and elovl5, key enzymes of LC-PUFA biosynthesis. Inhibition of pparγ 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 pparγ. 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 pparγ 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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References

  • Adeghate E, Adem A, Hasan MY, Tekes K, Kalasz H (2011) Suppl 2: medicinal chemistry and actions of dual and pan ppar modulators. Open Med Chem J 5:93–98

  • Baskerville S, Bartel DP (2005) Microarray profiling of microRNAs reveals frequent coexpression with neighboring miRNAs and host genes. RNA 11:241–247

    Article  CAS  Google Scholar 

  • Calder PC (2015) Very long chain omega-3 (n-3) fatty acids and human health. Eur J Lipid Sci Technol 116:1280–1300

    Article  Google Scholar 

  • Castro LF, Tocher DR, Monroig Ó (2016) Long-chain polyunsaturated fatty acid biosynthesis in chordates: insights into the evolution of fads and elovl gene repertoire. Prog Lipid Res 62:25–40

    Article  CAS  Google Scholar 

  • Chen CY, Sun BL, Guan WT, Bi YZ, Li PY, Ma J, Chen F, Pan Q, Xie QM (2016) N-3 essential fatty acids in Nile tilapia, Oreochromis niloticus: effects of linolenic acid on non-specific immunity and anti-inflammatory responses in juvenile fish. Aquaculture 450:250–257

    Article  CAS  Google Scholar 

  • Chen CY, Zhang JY, Zhang M, You CH, Liu Y, Wang SQ, Li YY (2018) miR-146a is involved in the regulation of vertebrate LC-PUFA biosynthesis by targeting elovl5 as demonstrated in rabbitfish siganus canaliculatus. Gene 676:306–314

  • Chen CY, Wang SQ, Zhang M, Chen BJ, You CH, Xie DZ, Liu Y, Zhang QH, Zhang JY, Monroig Ó, Tocher DR, Waiho K, Li YY (2019) miR-24 is involved in vertebrate LC-PUFA biosynthesis as demonstrated in marine teleost siganus canaliculatus. BBA-Mol Cell Biol L 1864:619

    CAS  Google Scholar 

  • Dong PY, Mai Y, Zhang ZY, Mi L, Wu GF, Chu GY, Yang GS, Sun SD (2014) miR-15a/b promote adipogenesis in porcine pre-adipocyte via repressing foxo1. Acta Biochim Biophys Sin 46:565–571

    Article  CAS  Google Scholar 

  • Dong YW, Wang SQ, Chen JL, Zhang QH, Liu Y, You CH, Monroig Ó, Tocher DR, Li YY (2016) Hepatocyte nuclear factor 4α (HNF4α) is a transcription factor of vertebrate fatty acyl desaturase gene as identified in marine teleost Siganus canaliculatus. PLoS One 11:e0160361

    Article  Google Scholar 

  • Dong YW, Zhao JH, Chen JL, Wang SQ, Liu Y, Zhang QH, You CH, Monroig Ó, Tocher DR, Li YY (2018) Cloning and characterization of ∆6/∆5 fatty acyl desaturase (fad) gene promoter in the marine teleost Siganus canaliculatus. Gene 647:174–180

    Article  CAS  Google Scholar 

  • Heikkinen S, Auwerx J, Argmann CA (2007) PPAR gamma in human and mouse physiology. Biochim Biophys Acta 1771:999–1013

    Article  CAS  Google Scholar 

  • Her GM, Hsu CC, Hong JR, Lai CY, Hsu MC, Pang HW, Chan SK, Pai WY (2011) Overexpression of gankyrin induces liver steatosis in zebrafish (danio rerio). BBA-Mol Cell Biol L 1811:536–548

    CAS  Google Scholar 

  • Janaki Ramaiah M, Lavanya A, Honarpisheh M, Zarea M, Bhadra U, Bhadra MP (2014) miR-15/16 complex targets p70s6 kinase1 and controls cell proliferation in mda-mb-231 breast cancer cells. Gene 552:255–264

    Article  CAS  Google Scholar 

  • Janssen CI, Kiliaan AJ (2014) Long-chain polyunsaturated fatty acids (LC-PUFA) from genesis to senescence: the influence of LC-PUFA on neural development, aging, and neurodegeneration. Prog Lipid Res 53:1–17

    Article  CAS  Google Scholar 

  • Lagos-Quintana M, Rauhut R, Lendeckel W (2001) Identification of novel genes coding for small expressed RNAs. Science 294:853–858

    Article  CAS  Google Scholar 

  • Li YY, Hu CB, Zheng YJ, Xia XA, Xu WJ, Wang SQ, Chen WZ, Sun ZW, Huang JH (2008) The effects of dietary fatty acids on liver fatty acid composition and Δ 6 -desaturase expression differ with ambient salinities in Siganus canaliculatus. Comp Biochem Physiol B 151:183–190

    Article  Google Scholar 

  • Li YY, Monroig Ó, Zhang L, Wang SQ, Zheng XZ, Dick JR, You CH, Tocher DR (2010) Vertebrate fatty acyl desaturase with Δ4 activity. Proc Natl Acad Sci USA 107:16840–16845

    Article  CAS  Google Scholar 

  • Li YY, Yin ZY, Dong YW, Wang SQ, Tocher DR, You CH (2019) Pparγ is involved in the transcriptional regulation of liver LC-PUFA biosynthesis by targeting the Δ6Δ5 fatty acyl desaturase gene in the marine teleost siganus canaliculatus. Mar Biotechnol 21:19–29

    Article  CAS  Google Scholar 

  • Liu Y, Zhang QH, Dong YW, You CH, Wang SQ, Li YQ, Li YY (2017) Establishment of a hepatocyte line for studying biosynthesis of long-chain polyunsaturated fatty acids from a marine teleost, the white-spotted spinefoot Siganus canaliculatus. J Fish Biol 91:603–616

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2012) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  Google Scholar 

  • Mangelsdorf DJ, Thummel C, Beato M, Herrlich P, Schütz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P, Evans RM (2000) The nuclear receptor superfamily: the second decade. Cell 83:835–839

    Article  Google Scholar 

  • Monroig Ó, Wang SQ, Zhang L, You CH, Tocher DR, Li YY (2012) Elongation of long-chain fatty acids in rabbitfish siganus canaliculatus: cloning, functional characterisation and tissue distribution of elovl5- and elovl4-like elongases. Aquaculture 350:63–70

    Article  Google Scholar 

  • Nordøy A, Dyerberg J (2015) N-3 fatty acids in health and disease. J Intern Med 225:1–3

    Article  Google Scholar 

  • Ou J, Tu H, Shan B, Luk A, Deboseboyd RA, Bashmakov Y, Goldstein JL, Brown MS (2001) Unsaturated fatty acids inhibit transcription of the sterol regulatory element-binding protein-1c (SREBP-1c) gene by antagonizing ligand-dependent activation of the LXR. Proc Natl Acad Sci U S A 98:6027–6032

    Article  CAS  Google Scholar 

  • Poulsen LLC, Siersbæk M, Mandrup S (2012) Ppars: fatty acid sensors controlling metabolism. Semin Cell Dev Biol 23:631–639

    Article  Google Scholar 

  • Poy MN, Eliasson L, Krutzfeldt J, Kuwajima J, Ma XS, Macdonald PE, Pfeffer S, Tuschl T, Rajewsky N, Rorsman P, Stoffel M (2004) A pancreatic islet-specific microRNA regulates insulin secretion. Nature 432:226–230

  • Sun JJ, Zheng LG, Chen CY, Zhang JY, You CH, Zhang QH, Ma HY, Monroig Ó, Tocher DR, Wang SQ, Li YY (2019) MicroRNAs involved in the regulation of LC-PUFA biosynthesis in teleosts: miR-33 enhances LC-PUFA biosynthesis in Siganus canaliculatus by targeting insig1 which in turn up-regulates srebp1. Mar Biotechnol 21:475–487

    Article  CAS  Google Scholar 

  • Tocher DR (2015) Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective. Aquaculture 449:94–107

    Article  CAS  Google Scholar 

  • Tocher DR, Bell JG, Dick JR, Crampton VO (2003) Effects of dietary vegetable oil on Atlantic salmon hepatocyte fatty acid desaturation and liver fatty acid compositions. Lipids 38:723–732

    Article  CAS  Google Scholar 

  • Tur JA, Bibiloni MM, Sureda A, Pons A (2012) Dietary sources of omega 3 fatty acids: public health risks and benefits. Br J Nutr 107:S23

    Article  CAS  Google Scholar 

  • Wang SQ, Chen JL, Jiang DL, Zhang QH, You CH, Tocher DR, Monroig Ó, Dong YW, Li YY (2018) Hnf4α is involved in the regulation of vertebrate LC-PUFA biosynthesis: insights into the regulatory role of Hnf4α on expression of liver fatty acyl desaturases in the marine teleost Siganus canaliculatus. Fish Physiol Biochem 44:1–11

    Article  Google Scholar 

  • You CH, Jiang DL, Zhang QH, Xie DZ, Wang SQ, Dong YW, Li YY (2017) Cloning and expression characterization of peroxisome proliferator-activated receptors (PPARs) with their agonists, dietary lipids, and ambient salinity in rabbitfish Siganus canaliculatus. Comp Biochem Physiol B 206:54–64

    Article  CAS  Google Scholar 

  • Yu J, Wang F, Yang GH, Wang FL, Ma YN, Du ZW, Zhang ZW (2006) Human microRNA clusters: genomic organization and expression profile in leukemia cell lines. Biochem Biophys Res Commun 349:59–68

    Article  CAS  Google Scholar 

  • Zhang QH, Xie DZ, Wang SQ, You CH, Monroig Ó, Tocher DR, Li YY (2014) miR-17 is involved in the regulation of LC-PUFA biosynthesis in vertebrates: effects on liver expression of a fatty acyl desaturase in the marine teleost Siganus canaliculatus. BBA-Mol Cell Biol L 1841:934–943

    CAS  Google Scholar 

  • Zhang QH, You CH, Liu F, Zhu WD, Wang SQ, Xie DZ, Monroig Ó, Tocher DR, Li YY (2016a) Cloning and characterization of lxr and srebp1, and their potential roles in regulation of LC-PUFA biosynthesis in rabbitfish siganus canaliculatus. Lipids 51:1051–1063

    Article  CAS  Google Scholar 

  • Zhang QH, You CH, Wang SQ, Dong YW, Monroig Ó, Tocher DR, Li YY (2016b) The miR-33 gene is identified in a marine teleost: a potential role in regulation of LC-PUFA biosynthesis in Siganus canaliculatus. Sci Rep 6:32909

    Article  CAS  Google Scholar 

Download references

Funding

This work was financially supported by the National Key R&D Program of China (2018YFD0900400), National Natural Science Foundation of China (No. 31873040 & No. 31702357), Natural Science Foundation of Guangdong Province (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), and Natural Science Foundation of Guangdong Province (2020A1515011252).

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Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Marine Biotechnology, Institute of Marine Sciences, Shantou University, Shantou, 515063, China

    Junjun Sun, Cuiying Chen, Cuihong You, Yang Liu, Hongyu Ma & Shuqi Wang

  2. School of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China

    Yuanyou Li

  3. STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China

    Junjun Sun, Cuiying Chen, Cuihong You, Hongyu Ma & Shuqi Wang

  4. Instituto de Acuicultura Torre de la Sal, Consejo Superior de Investigaciones Científicas (IATS-CSIC), 12595 Ribera de Cabanes, Castellón, Spain

    Óscar Monroig

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

    Douglas R. Tocher

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  1. Junjun Sun
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  2. Cuiying Chen
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Correspondence to Shuqi Wang or Yuanyou Li.

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Sun, J., Chen, C., You, C. et al. The miR-15/16 Cluster Is Involved in the Regulation of Vertebrate LC-PUFA Biosynthesis by Targeting pparγ as Demonostrated in Rabbitfish Siganus canaliculatus. Mar Biotechnol 22, 475–487 (2020). https://doi.org/10.1007/s10126-020-09969-0

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