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Characterization of an oleate 12-desaturase from Physaria fendleri and identification of 5'UTR introns in divergent FAD2 family genes.
Plant Physiol Biochem. 2014 Feb; 75:114-22.PP

Abstract

Mining of an EST sequence collection representing genes expressed during seed development in Physaria fendleri identified abundant sequences encoding apparent homologues of the Arabidopsis oleate 12-desaturase (AtFAD2 At3g12120). Of the 62 sequenced clones, 59 were identified as encoding the previously characterized bifunctional oleate 12-hydroxylase/desaturase (LFAH12/PfFAH12). The remaining 3 clones encoded a second FAD2 homologue. Isolation of a full length ORF and heterologous expression in yeast revealed that this sequence, designated PfFAD2, is the first full length sequence from any Physaria species that encodes an oleate 12-desaturase. PfFAD2 was expressed in both leaf and developing seed with activity on palmitate (16:1(Δ9)) and oleate (18:1(Δ9)). Sequence comparison revealed that PfFAD2 shares 93% amino acid identity with Arabidopsis FAD2 and only 84% identity with PfFAH12. By comparison of EST and genomic sequences it was revealed that the PfFAD2 gene encodes a transcript with a single intron of 1120 bp in the 5'-untranslated region (5'UTR). A short intron, 81 bp in length, was also discovered in the 5'UTR of the PfFAH12 gene, 16 bp upstream of the translation initiation codon. In silico examination of FAD2 like genes from the genome of castor (Ricinus communis) identified putative 5'UTR introns in genes encoding the castor oleate 12-desaturase (RcFAD2) and oleate 12-hydroxylase (CFAH12). By sequencing of genomic DNA the presence of single 5'UTR introns in each gene, and the size of these introns, was confirmed. These findings suggest that 5'UTR introns may be a characteristic feature of FAD2 genes and also of divergent FAD2 genes encoding fatty acid modifying enzymes, and that the selection pressure maintaining these introns is very different.

Authors+Show Affiliations

National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada. Electronic address: Mark.Smith@nrc-cnrc.gc.ca.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24429134

Citation

Lozinsky, Sharla, et al. "Characterization of an Oleate 12-desaturase From Physaria Fendleri and Identification of 5'UTR Introns in Divergent FAD2 Family Genes." Plant Physiology and Biochemistry : PPB, vol. 75, 2014, pp. 114-22.
Lozinsky S, Yang H, Forseille L, et al. Characterization of an oleate 12-desaturase from Physaria fendleri and identification of 5'UTR introns in divergent FAD2 family genes. Plant Physiol Biochem. 2014;75:114-22.
Lozinsky, S., Yang, H., Forseille, L., Cook, G. R., Ramirez-Erosa, I., & Smith, M. A. (2014). Characterization of an oleate 12-desaturase from Physaria fendleri and identification of 5'UTR introns in divergent FAD2 family genes. Plant Physiology and Biochemistry : PPB, 75, 114-22. https://doi.org/10.1016/j.plaphy.2013.12.016
Lozinsky S, et al. Characterization of an Oleate 12-desaturase From Physaria Fendleri and Identification of 5'UTR Introns in Divergent FAD2 Family Genes. Plant Physiol Biochem. 2014;75:114-22. PubMed PMID: 24429134.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of an oleate 12-desaturase from Physaria fendleri and identification of 5'UTR introns in divergent FAD2 family genes. AU - Lozinsky,Sharla, AU - Yang,Hui, AU - Forseille,Li, AU - Cook,Gillian R, AU - Ramirez-Erosa,Irving, AU - Smith,Mark A, Y1 - 2013/12/28/ PY - 2013/10/30/received PY - 2013/12/20/accepted PY - 2014/1/17/entrez PY - 2014/1/17/pubmed PY - 2014/9/30/medline KW - 5′UTR intron KW - EST KW - FAD2 KW - FAMES KW - Fatty acid desaturase KW - Fatty acid hydroxylase KW - HFA KW - Lesquerella KW - ORF KW - Physaria fendleri KW - expressed sequence tag KW - fatty acid desaturase 2 KW - fatty acid methyl esters KW - hydroxy fatty acid KW - open reading frame SP - 114 EP - 22 JF - Plant physiology and biochemistry : PPB JO - Plant Physiol Biochem VL - 75 N2 - Mining of an EST sequence collection representing genes expressed during seed development in Physaria fendleri identified abundant sequences encoding apparent homologues of the Arabidopsis oleate 12-desaturase (AtFAD2 At3g12120). Of the 62 sequenced clones, 59 were identified as encoding the previously characterized bifunctional oleate 12-hydroxylase/desaturase (LFAH12/PfFAH12). The remaining 3 clones encoded a second FAD2 homologue. Isolation of a full length ORF and heterologous expression in yeast revealed that this sequence, designated PfFAD2, is the first full length sequence from any Physaria species that encodes an oleate 12-desaturase. PfFAD2 was expressed in both leaf and developing seed with activity on palmitate (16:1(Δ9)) and oleate (18:1(Δ9)). Sequence comparison revealed that PfFAD2 shares 93% amino acid identity with Arabidopsis FAD2 and only 84% identity with PfFAH12. By comparison of EST and genomic sequences it was revealed that the PfFAD2 gene encodes a transcript with a single intron of 1120 bp in the 5'-untranslated region (5'UTR). A short intron, 81 bp in length, was also discovered in the 5'UTR of the PfFAH12 gene, 16 bp upstream of the translation initiation codon. In silico examination of FAD2 like genes from the genome of castor (Ricinus communis) identified putative 5'UTR introns in genes encoding the castor oleate 12-desaturase (RcFAD2) and oleate 12-hydroxylase (CFAH12). By sequencing of genomic DNA the presence of single 5'UTR introns in each gene, and the size of these introns, was confirmed. These findings suggest that 5'UTR introns may be a characteristic feature of FAD2 genes and also of divergent FAD2 genes encoding fatty acid modifying enzymes, and that the selection pressure maintaining these introns is very different. SN - 1873-2690 UR - https://www.unboundmedicine.com/medline/citation/24429134/Characterization_of_an_oleate_12_desaturase_from_Physaria_fendleri_and_identification_of_5'UTR_introns_in_divergent_FAD2_family_genes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0981-9428(13)00440-3 DB - PRIME DP - Unbound Medicine ER -