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Molecular Biology Reports

, Volume 43, Issue 8, pp 761–766 | Cite as

Alternative splicing generates novel Fads3 transcript in mice

  • Ji Yao Zhang
  • Xia Qin
  • Hui Gyu Park
  • Ellen Kim
  • Guowen Liu
  • Kumar S. D. KothapalliEmail author
  • J. Thomas BrennaEmail author
Short Communication

Abstract

Fads3 is the third member of the fatty acid desaturase gene cluster; with at least eight evolutionarily conserved alternative transcripts (AT), having no clearly established function as are known for FADS2 and FADS1. Here we present identification of a novel Fads3 transcript in mice (Fads3AT9), characterize Fads3AT9 expression in mouse tissues and evaluate correlations with metabolite profiles. Total RNA obtained from mouse tissues is reverse-transcribed into cDNA and used as template for PCR reactions. Tissue fatty acids were extracted and quantified by gas chromatography. Sequencing analysis revealed complete absence of exon 2 resulting in an open reading frame of 1239 bp, encoding a putative protein of 412 aa with loss of 37 aa compared to classical Fads3 (Fads3CS). FADS3AT9 retains all the conserved regions characteristic of front end desaturase (cytochrome b5 domain and three histidine repeats). Both Fads3CS and Fads3AT9 are ubiquitously expressed in 11 mouse tissues. Fads3AT9 abundance was greater than Fads3CS in pancreas, liver, spleen, brown adipose tissue and thymus. Fads3CS expression is low in pancreas while Fads3AT9 is over ten-fold greater abundance. The eicosanoid precursor fatty acid 20:4n − 6, the immediate desaturation product of the Fads1 coded Δ5-desaturase, was highest in pancreas where Fads3CS is low. Changes in expression patterns and fatty acid profiles suggest that Fads3AT9 may play a role in the regulation and/or biosynthesis of long chain polyunsaturated fatty acids from precursors.

Keywords

Fatty acid desaturase Alternative transcript Alternative splicing Long chain polyunsaturated fatty acids 

Notes

Acknowledgments

This work was supported by NIH grant R01 AT007003 from the National Center for Complementary and Integrative Health (NCCIH) and the Office of Dietary Supplements (ODS). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.

Author Contributions

J.T.B., K.S.D.K., X.Q., J.Y.Z. designed research; X.Q., J.Y.Z., H.P., and E.K. executed the research; J.T. B., K.S.D.K., and G.L. contributed new reagents/analytic tools; J.T.B., K.S.D.K., X.Q., and J.Y.Z. analyzed and interpreted the data; and J.T. B., K.S.D.K., X.Q., and J.Y.Z. wrote the first draft and all authors approved the final draft.

Compliance with ethical standards

Conflict of interest

All authors declare no conflict of interest.

Supplementary material

11033_2016_4018_MOESM1_ESM.docx (548 kb)
Supplementary material 1 (DOCX 547 kb)

References

  1. 1.
    Park WJ et al (2011) FADS2 function loss at the cancer hotspot 11q13 locus diverts lipid signaling precursor synthesis to unusual eicosanoid fatty acids. Plos One 6(11):e28186CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Park HG et al (2015) The fatty acid desaturase 2 (FADS2) gene product catalyzes Delta4 desaturation to yield n-3 docosahexaenoic acid and n-6 docosapentaenoic acid in human cells. FASEB J 29(9):3911–3919CrossRefPubMedGoogle Scholar
  3. 3.
    Marquardt A et al (2000) cDNA cloning, genomic structure, and chromosomal localization of three members of the human fatty acid desaturase family. Genomics 66(2):175–183CrossRefPubMedGoogle Scholar
  4. 4.
    Nakamura MT, Nara TY (2004) Structure, function, and dietary regulation of delta6, delta5, and delta9 desaturases. Annu Rev Nutr 24:345–376CrossRefPubMedGoogle Scholar
  5. 5.
    Park WJ et al (2012) A novel FADS1 isoform potentiates FADS2-mediated production of eicosanoid precursor fatty acids. J Lipid Res 53(8):1502–1512CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Ma XH et al (2006) Serial analysis of gene expression in mouse uterus at the implantation site. J Biol Chem 281(14):9351–9360CrossRefPubMedGoogle Scholar
  7. 7.
    Zhang J et al (2014) Fatty acid desaturase 3 (Fads3) null mouse biochemical phenotype. FASEB J 28(1 Supplement):246Google Scholar
  8. 8.
    Rioux V et al (2013) Trans-vaccenate is Delta13-desaturated by FADS3 in rodents. J Lipid Res 54(12):3438–3452CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Plaisier CL et al (2009) A systems genetics approach implicates USF1, FADS3, and other causal candidate genes for familial combined hyperlipidemia. PLoS Genet 5(9):e1000642CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Koletzko B et al (2011) Genetic variants of the fatty acid desaturase gene cluster predict amounts of red blood cell docosahexaenoic and other polyunsaturated fatty acids in pregnant women: findings from the avon longitudinal study of parents and children. Am J Clin Nutr 93(1):211–219CrossRefPubMedGoogle Scholar
  11. 11.
    Park WJ et al (2009) Novel fatty acid desaturase 3 (FADS3) transcripts generated by alternative splicing. Gene 446(1):28–34CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Pedrono F et al (2010) The fatty acid desaturase 3 gene encodes for different FADS3 protein isoforms in mammalian tissues. J Lipid Res 51(3):472–479CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Park WJ et al (2010) Alternative splicing generates a novel FADS2 alternative transcript in baboons. Mol Biol Rep 37(5):2403–2406CrossRefPubMedGoogle Scholar
  14. 14.
    Garces R, Mancha M (1993) One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal Biochem 211(1):139–143CrossRefPubMedGoogle Scholar
  15. 15.
    Chou KC, Shen HB (2008) Cell-PLoc: a package of Web servers for predicting subcellular localization of proteins in various organisms. Nat Protoc 3(2):153–162CrossRefPubMedGoogle Scholar
  16. 16.
    Shoji Y et al (2003) Preparation of antiserum against rat delta6-desaturase and its use to evaluate the desaturase protein levels in rats treated with gemfibrozil, a ligand for peroxisome proliferator-activated receptor alpha. Biosci Biotechnol Biochem 67(5):1177–1178CrossRefPubMedGoogle Scholar
  17. 17.
    Pinnick KE et al (2008) Pancreatic ectopic fat is characterized by adipocyte infiltration and altered lipid composition. Obesity (Silver Spring) 16(3):522–530CrossRefGoogle Scholar
  18. 18.
    Kelemen O et al (2013) Function of alternative splicing. Gene 514(1):1–30CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Ji Yao Zhang
    • 1
  • Xia Qin
    • 1
    • 2
  • Hui Gyu Park
    • 1
  • Ellen Kim
    • 1
  • Guowen Liu
    • 2
  • Kumar S. D. Kothapalli
    • 1
    Email author
  • J. Thomas Brenna
    • 1
    Email author
  1. 1.Division of Nutritional SciencesCornell UniversityIthacaUSA
  2. 2.College of Veterinary MedicineJilin UniversityJilinChina

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