A vertebrate fatty acid desaturase with Δ5 and Δ6 activities

Hastings, Nicola; Agaba, Morris; Tocher, Douglas R.; Leaver, Michael J.; Dick, James R.; Sargent, J. R.; Teale, A.J.

doi:10.1073/pnas.251516598

Cited by 311 publications

(303 citation statements)

References 43 publications

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“…Gene duplication events in mammals have resulted in multiple gene copies encoding desaturase (Fads1, 2, and 3) enzymes, whereas in other vertebrates (such as Danio rerio ), only a single desaturase, with Δ5 and Δ6 activity, is present (Hastings et al., 2001). Similarly, whole genome duplication events have resulted in the diversification of elongase enzymes observed in vertebrates (Elovl2 and 5) not present in other chordates, which contain a single elongase enzyme referred to as Elovl2/5 (Castro, Tocher, & Monroig, 2016; Monroig et al., 2016).…”

Section: Introductionmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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The biosynthesis of long‐chain polyunsaturated fatty acids (LC‐PUFAs, ≥ C20) is reliant on the action of desaturase and elongase enzymes, which are encoded by the fatty acyl desaturase (Fad) and elongation of very long‐chain fatty acid (Elovl) gene families, respectively. In Metazoa, research investigating the distribution and evolution of these gene families has been restricted largely to Bilateria. Here, we provide insights into the phylogenetic and molecular evolutionary histories of the Fad and Elovl gene families in Cnidaria, the sister phylum to Bilateria. Four model cnidarian genomes and six actiniarian transcriptomes were interrogated. Analysis of the fatty acid composition of a candidate cnidarian species, Actinia tenebrosa, was performed to determine the baseline profile of this species. Phylogenetic analysis revealed lineage‐specific gene duplication in actiniarians for both the Fad and Elovl gene families. Two distinct cnidarian Fad clades clustered with functionally characterized Δ5 and Δ6 proteins from fungal and plant species, respectively. Alternatively, only a single cnidarian Elovl clade clustered with functionally characterized Elovl proteins (Elovl4), while two additional clades were identified, one actiniarian‐specific (Novel ElovlA) and the another cnidarian‐specific (Novel ElovlB). In actiniarians, selection analyses revealed pervasive purifying selection acting on both gene families. However, codons in the Elovl gene family show patterns of nucleotide variation consistent with the action of episodic diversifying selection following gene duplication events. Significantly, these codons may encode amino acid residues that are functionally important for Elovl proteins to target and elongate different precursor fatty acids. In A. tenebrosa, the fatty acid analysis revealed an absence of LC‐PUFAs > C20 molecules and implies that the Elovl enzymes are not actively contributing to the elongation of these LC‐PUFAs. Overall, this study has revealed that actiniarians possess Fad and Elovl genes required for the biosynthesis of some LC‐PUFAs, and that these genes appear to be distinct from bilaterians.

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Section: Introductionmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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“…Fatty acid Δ5 desaturase genes have been isolated from M. alpina (Michaelson et al, 1998a), C. elegans (Michaelson et al, 1998b;Watts and Browse, 1999) and human (Cho et al, 1999b;Leonard et al, 2000). Using extant sequence information, we isolated a cDNA from the freshwater zebrafish (Danio rerio) with high similarity to mammalian Δ6 desaturase genes (Hastings et al, 2001). Heterologous expression in yeast indicated that the zebrafish desaturase gene was unique in that it encoded an enzyme having both Δ6 and Δ5 desaturase activities, prompting us to speculate that it may represent a component of a prototypic vertebrate HUFA biosynthesis pathway.…”

Section: Introductionmentioning

confidence: 99%

“…The RNA extraction, cDNA synthesis, primers and PCR conditions for cloning putative PUFA desaturase from carp (GenBank accession no. AF309557) were coincidentally the same as for the zebrafish desaturase as described in detail previously (Hastings et al, 2001).…”

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confidence: 99%

“…Following PCR, the DNA fragments were restricted with the appropriate enzymes, HindIII and Culture of the recombinant yeast was carried out in SCMM -uracil broth using galactose induction of gene expression as described previously (Hastings et al, 2001). Cultures were supplemented with one of the PUFA substrates from among the following; α-linolenic acid (18:3n-3), linoleic acid (18:2n-6), eicosatetraenoic acid (20:4n-3), dihomo-γ-linolenic acid (20:3n-6), docosapentaenoic acid (22:5n-3) and docosatetraenoic acid (22:4n-6), Yeast cells were harvested, washed and dried under oxygen-free nitrogen as described previously (Hastings et al, 2001), Lipid was extracted by homogenisation in chloroform/methanol (2:1) containing 0.01% butylated hydroxytoluene (BHT) as antioxidant, essentially according to Folch et al (1957), and fatty acid methyl esters (FAME) prepared by addition of 1 ml toluene and 2 ml 1% methanolic sulphuric acid and incubation overnight at 50 o C (Christie 1982).…”

mentioning

confidence: 99%

“…Cultures were supplemented with one of the PUFA substrates from among the following; α-linolenic acid (18:3n-3), linoleic acid (18:2n-6), eicosatetraenoic acid (20:4n-3), dihomo-γ-linolenic acid (20:3n-6), docosapentaenoic acid (22:5n-3) and docosatetraenoic acid (22:4n-6), Yeast cells were harvested, washed and dried under oxygen-free nitrogen as described previously (Hastings et al, 2001), Lipid was extracted by homogenisation in chloroform/methanol (2:1) containing 0.01% butylated hydroxytoluene (BHT) as antioxidant, essentially according to Folch et al (1957), and fatty acid methyl esters (FAME) prepared by addition of 1 ml toluene and 2 ml 1% methanolic sulphuric acid and incubation overnight at 50 o C (Christie 1982). The FAME were extracted, purified by thin layer chromatography (TLC), and analysed by gas chromatography (GC) all as described previously 7 (Hastings et al, 2001). The proportion of substrate fatty acid converted to the desaturated fatty acid product was calculated from the gas chromatograms as 100 × [product area/(product area + substrate area)].…”

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confidence: 99%

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Characterization and comparison of fatty acyl Δ6 desaturase cDNAs from freshwater and marine teleost fish species

Zheng

Seiliez

Hastings

et al. 2004

Comparative Biochemistry and Physiology Part B: Biochemistry an

173

139

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Abstract:Fish are the most important dietary source of the n-3 highly unsaturated fatty acids (HUFA), eicosapentaenoic (EPA) and docosahexaenoic acid (DHA), that have particularly important roles in human nutrition reflecting their roles in critical physiological processes. The objective of the study described here was to clone, functionally characterise and compare expressed fatty acid desaturase genes involved in the production of EPA and DHA in freshwater and marine teleost fish species.Putative fatty acid desaturase cDNAs were isolated and cloned from common carp (Cyprinus carpio) and turbot (Psetta maximus). The enzymic activities of the products of these cDNAs, together with those of cDNAs previously cloned from rainbow trout (Oncorhynchus mykiss) and gilthead seabream (Sparus aurata), were determined by heterologous expression in the yeast containing the haem-binding motif, HPGG. Functional expression showed all four fish cDNAs encode basically unifunctional Δ6 fatty acid desaturase enzymes responsible for the first and ratelimiting step in the biosynthesis of HUFA from 18:3n-3 and 18:2n-6. All the fish desaturases were more active towards the n-3 substrate with 59.5%, 31.5%, 23.1% and 7.0% of 18:3n-3 being converted to 18:4n-3 in the case of turbot, trout, seabream and carp, respectively. The enzymes also showed very low, probably physiologically insignificant, levels of Δ5 desaturase activity, but none of the products showed Δ4 desaturase activity. The cloning and characterisation of desaturases from these fish is an important advance, as they are species in which there is a relative wealth of data on the nutritional regulation of fatty acid desaturation and HUFA synthesis, and between which substantive differences occur.

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Genome‐wide comparative identification and analysis of membrane‐FADS‐like superfamily genes in freshwater economic fishes

Zhang

Gao

et al. 2023

FEBS Open Bio

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Membrane fatty acid desaturase (FADS)‐like superfamily proteins (FADSs) are essential for the synthesis of unsaturated fatty acids (UFAs). Recently, studies on FADS in fishes have mostly focused on marine species, and a comprehensive analysis of the FADS superfamily, including the FADS, stearoyl‐CoA desaturase (SCD), and sphingolipid delta 4‐desaturase (DEGS) families, in freshwater economic fishes is urgently required. To this end, we conducted a thorough analysis of the number, gene/protein structure, chromosomal location, gene linkage map, phylogeny, and expression of the FADS superfamily. We identified 156 FADSs genes in the genome of 27 representative species. Notably, FADS1 and SCD5 were lost in most freshwater fish and other teleosts. All FADSs proteins contain 4 transmembrane helices and 2–3 amphipathic α‐helices. FADSs in the same family are often linked on the same chromosome; moreover, FADS and SCD or DEGS are frequently collocated on the same chromosome. In addition, FADS, SCD, and DEGS family proteins share similar evolutionary patterns. Interestingly, FADS6, as a member of the FADS family, exhibits a similar gene structure and chromosome location to that of SCD family members, which may be the transitional form of FADS and SCD. This study shed light on the type, structure, and phylogenetic relationship of FADSs in freshwater fishes, offering a new perspective into the functional mechanism analysis of FADSs.

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A vertebrate fatty acid desaturase with Δ5 and Δ6 activities

Cited by 311 publications

References 43 publications

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Characterization and comparison of fatty acyl Δ6 desaturase cDNAs from freshwater and marine teleost fish species

Genome‐wide comparative identification and analysis of membrane‐FADS‐like superfamily genes in freshwater economic fishes

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