Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 May 1;347(Pt 3):719–724.

cDNA cloning and characterization of human Delta5-desaturase involved in the biosynthesis of arachidonic acid.

A E Leonard 1, B Kelder 1, E G Bobik 1, L T Chuang 1, J M Parker-Barnes 1, J M Thurmond 1, P E Kroeger 1, J J Kopchick 1, Y S Huang 1, P Mukerji 1
PMCID: PMC1221008  PMID: 10769175

Abstract

Two human expressed sequence tag (EST) cDNA sequences with identity with Delta(5)- and Delta(6)-desaturases from a filamentous fungus, Mortierella alpina, were identified from the LifeSeq(R) database of Incyte Pharmaceuticals, Inc. (Palo Alto, CA, U.S.A.). An oligonucleotide complementary to the 3' EST cDNA sequences was used to screen human liver cDNA using rapid amplification of cDNA ends (RACE)-PCR. The amplified DNA fragment had 98% identity with a putative open reading frame (ORF) predicted from a human genomic sequence, and encoded 444 amino acids. Expression of this ORF in mouse fibroblast cells demonstrated that the encoded protein was a Delta(5)-desaturase, as determined by the conversion of dihomo-gamma-linolenic acid (C(20:3,n-6)) into arachidonic acid (C(20:4,n-6)). The human Delta(5)-desaturase contained a predicted N-terminal cytochrome b(5)-like domain, as well as three histidine-rich domains. A tissue expression profile revealed that this gene is highly expressed in fetal liver, fetal brain, adult brain and adrenal gland. A search of the existing databases led to localization of this ORF within a 14 kb interval flanked by the flap endonuclease-1 (FEN1) and vitelliform macular dystrophy (Best's disease; VMD2) loci of chromosome 11q12.

Full Text

The Full Text of this article is available as a PDF (330.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguilar P. S., Cronan J. E., Jr, de Mendoza D. A Bacillus subtilis gene induced by cold shock encodes a membrane phospholipid desaturase. J Bacteriol. 1998 Apr;180(8):2194–2200. doi: 10.1128/jb.180.8.2194-2200.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aki T., Shimada Y., Inagaki K., Higashimoto H., Kawamoto S., Shigeta S., Ono K., Suzuki O. Molecular cloning and functional characterization of rat delta-6 fatty acid desaturase. Biochem Biophys Res Commun. 1999 Feb 24;255(3):575–579. doi: 10.1006/bbrc.1999.0235. [DOI] [PubMed] [Google Scholar]
  3. Birch E. E., Hoffman D. R., Uauy R., Birch D. G., Prestidge C. Visual acuity and the essentiality of docosahexaenoic acid and arachidonic acid in the diet of term infants. Pediatr Res. 1998 Aug;44(2):201–209. doi: 10.1203/00006450-199808000-00011. [DOI] [PubMed] [Google Scholar]
  4. Carlson S. E., Werkman S. H., Peeples J. M., Cooke R. J., Tolley E. A. Arachidonic acid status correlates with first year growth in preterm infants. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1073–1077. doi: 10.1073/pnas.90.3.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cho H. P., Nakamura M. T., Clarke S. D. Cloning, expression, and nutritional regulation of the mammalian Delta-6 desaturase. J Biol Chem. 1999 Jan 1;274(1):471–477. doi: 10.1074/jbc.274.1.471. [DOI] [PubMed] [Google Scholar]
  6. Girke T., Schmidt H., Zähringer U., Reski R., Heinz E. Identification of a novel delta 6-acyl-group desaturase by targeted gene disruption in Physcomitrella patens. Plant J. 1998 Jul;15(1):39–48. doi: 10.1046/j.1365-313x.1998.00178.x. [DOI] [PubMed] [Google Scholar]
  7. Gong Y., Vikkula M., Boon L., Liu J., Beighton P., Ramesar R., Peltonen L., Somer H., Hirose T., Dallapiccola B. Osteoporosis-pseudoglioma syndrome, a disorder affecting skeletal strength and vision, is assigned to chromosome region 11q12-13. Am J Hum Genet. 1996 Jul;59(1):146–151. [PMC free article] [PubMed] [Google Scholar]
  8. Horrobin D. F. Nutritional and medical importance of gamma-linolenic acid. Prog Lipid Res. 1992;31(2):163–194. doi: 10.1016/0163-7827(92)90008-7. [DOI] [PubMed] [Google Scholar]
  9. Huang Y. S., Chaudhary S., Thurmond J. M., Bobik E. G., Jr, Yuan L., Chan G. M., Kirchner S. J., Mukerji P., Knutzon D. S. Cloning of delta12- and delta6-desaturases from Mortierella alpina and recombinant production of gamma-linolenic acid in Saccharomyces cerevisiae. Lipids. 1999 Jul;34(7):649–659. doi: 10.1007/s11745-999-0410-8. [DOI] [PubMed] [Google Scholar]
  10. Kelder B., Richmond C., Stavnezer E., List E. O., Kopchick J. J. Production, characterization and functional activities of v-Ski in cultured cells. Gene. 1997 Nov 20;202(1-2):15–21. doi: 10.1016/s0378-1119(97)00439-3. [DOI] [PubMed] [Google Scholar]
  11. Knutzon D. S., Thurmond J. M., Huang Y. S., Chaudhary S., Bobik E. G., Jr, Chan G. M., Kirchner S. J., Mukerji P. Identification of Delta5-desaturase from Mortierella alpina by heterologous expression in Bakers' yeast and canola. J Biol Chem. 1998 Nov 6;273(45):29360–29366. doi: 10.1074/jbc.273.45.29360. [DOI] [PubMed] [Google Scholar]
  12. Leppert M., Baird L., Anderson K. L., Otterud B., Lupski J. R., Lewis R. A. Bardet-Biedl syndrome is linked to DNA markers on chromosome 11q and is genetically heterogeneous. Nat Genet. 1994 May;7(1):108–112. doi: 10.1038/ng0594-108. [DOI] [PubMed] [Google Scholar]
  13. Li Y., Müller B., Fuhrmann C., van Nouhuys C. E., Laqua H., Humphries P., Schwinger E., Gal A. The autosomal dominant familial exudative vitreoretinopathy locus maps on 11q and is closely linked to D11S533. Am J Hum Genet. 1992 Oct;51(4):749–754. [PMC free article] [PubMed] [Google Scholar]
  14. Marquardt A., Stöhr H., Passmore L. A., Krämer F., Rivera A., Weber B. H. Mutations in a novel gene, VMD2, encoding a protein of unknown properties cause juvenile-onset vitelliform macular dystrophy (Best's disease). Hum Mol Genet. 1998 Sep;7(9):1517–1525. doi: 10.1093/hmg/7.9.1517. [DOI] [PubMed] [Google Scholar]
  15. Napier J. A., Hey S. J., Lacey D. J., Shewry P. R. Identification of a Caenorhabditis elegans Delta6-fatty-acid-desaturase by heterologous expression in Saccharomyces cerevisiae. Biochem J. 1998 Mar 1;330(Pt 2):611–614. doi: 10.1042/bj3300611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Samuelsson B. From studies of biochemical mechanism to novel biological mediators: prostaglandin endoperoxides, thromboxanes, and leukotrienes. Nobel Lecture, 8 December 1982. Biosci Rep. 1983 Sep;3(9):791–813. doi: 10.1007/BF01133779. [DOI] [PubMed] [Google Scholar]
  17. Sayanova O., Smith M. A., Lapinskas P., Stobart A. K., Dobson G., Christie W. W., Shewry P. R., Napier J. A. Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of delta6-desaturated fatty acids in transgenic tobacco. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):4211–4216. doi: 10.1073/pnas.94.8.4211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stone E. M., Kimura A. E., Folk J. C., Bennett S. R., Nichols B. E., Streb L. M., Sheffield V. C. Genetic linkage of autosomal dominant neovascular inflammatory vitreoretinopathy to chromosome 11q13. Hum Mol Genet. 1992 Dec;1(9):685–689. doi: 10.1093/hmg/1.9.685. [DOI] [PubMed] [Google Scholar]
  19. Stöhr H., Marquardt A., Rivera A., Cooper P. R., Nowak N. J., Shows T. B., Gerhard D. S., Weber B. H. A gene map of the Best's vitelliform macular dystrophy region in chromosome 11q12-q13.1. Genome Res. 1998 Jan;8(1):48–56. doi: 10.1101/gr.8.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Uauy R., Birch E., Birch D., Peirano P. Visual and brain function measurements in studies of n-3 fatty acid requirements of infants. J Pediatr. 1992 Apr;120(4 Pt 2):S168–S180. doi: 10.1016/s0022-3476(05)81252-1. [DOI] [PubMed] [Google Scholar]
  21. Watts J. L., Browse J. Isolation and characterization of a Delta 5-fatty acid desaturase from Caenorhabditis elegans. Arch Biochem Biophys. 1999 Feb 1;362(1):175–182. doi: 10.1006/abbi.1998.1024. [DOI] [PubMed] [Google Scholar]
  22. Yoo M., Steggles A. W. The complete nucleotide sequence of human liver cytochrome b5 mRNA. Biochem Biophys Res Commun. 1988 Oct 14;156(1):576–580. doi: 10.1016/s0006-291x(88)80881-7. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES