Polyunsaturated fatty acids (PUFA) influence many physiological functions. Associations have been found between single nucleotide polymorphisms (SNP) in the FADS1 (Fatty acid desaturase 1) gene and the relative abundance of PUFA in serum lipids. This study examines the relationship between two SNPs in the FADS1 gene (rs174546, rs174537) and the fatty acid (FA) composition of serum lipids in adolescents (13-18 years). We used DNA samples (670 children; 336 girls and 334 boys) from the Childhood Obesity Prevalence and Treatment (COPAT) project. Genomic DNA was extracted from peripheral blood leukocytes in whole blood samples. For genotype analysis, TaqMan SNP Genotyping assays (Applied Biosystems) were used. Fatty acid composition of serum lipids was assessed using gas chromatography. The T-statistic and regression were used for statistical evaluations. Minor allele T carriers in both SNPs had significant lower level of palmitic acid (16:0, phospholipids) and arachidonic acid (20:4[n-6], phospholipids) in both sexes. In girls, we found a significant positive association between minor allele T carriers and eicosadienoic acid (20:2[n-6], cholesteryl esters) in both SNPs. Being a minor allele T carrier was significantly positively associated with dihomo-γ-linolenic acid (20:3[n-6], phospholipids) in boys in both SNPs. SNPs (including rs174546, rs174537) in the FADS gene cluster should have impacted desaturase activity, which may contribute to different efficiency of PUFA synthesis.
- MeSH
- alely MeSH
- delta-5 desaturasa mastných kyselin MeSH
- desaturasy mastných kyselin genetika MeSH
- dítě MeSH
- genotyp MeSH
- jednonukleotidový polymorfismus MeSH
- lidé MeSH
- mastné kyseliny * MeSH
- mladiství MeSH
- obezita dětí a dospívajících * MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The development of colon cancer, one of the most common malignancies, is accompanied with numerous lipid alterations. However, analyses of whole tumor samples may not always provide an accurate description of specific changes occurring directly in tumor epithelial cells. Here, we analyzed in detail the phospholipid (PL), lysophospholipid (lysoPL), and fatty acid (FA) profiles of purified EpCAM+ cells, isolated from tumor and adjacent non-tumor tissues of colon cancer patients. We found that a number of FAs increased significantly in isolated tumor cells, which also included a number of long polyunsaturated FAs. Higher levels of FAs were associated with increased expression of FA synthesis genes, as well as with altered expression of enzymes involved in FA elongation and desaturation, including particularly fatty acid synthase, stearoyl-CoA desaturase, fatty acid desaturase 2 and ELOVL5 fatty acid elongase 5 We identified significant changes in ratios of specific lysoPLs and corresponding PLs. A number of lysophosphatidylcholine and lysophosphatidylethanolamine species, containing long-chain and very-long chain FAs, often with high numbers of double bonds, were significantly upregulated in tumor cells. Increased de novo synthesis of very long-chain FAs, or, altered uptake or incorporation of these FAs into specific lysoPLs in tumor cells, may thus contribute to reprogramming of cellular phospholipidome and membrane alterations observed in colon cancer.
- MeSH
- adenokarcinom enzymologie genetika metabolismus MeSH
- desaturasy mastných kyselin genetika metabolismus MeSH
- elongasy mastných kyselin genetika metabolismus MeSH
- epitelové buňky enzymologie metabolismus MeSH
- fosfolipidy metabolismus MeSH
- lidé MeSH
- lipidomika MeSH
- lipogeneze MeSH
- mastné kyseliny metabolismus MeSH
- metabolismus lipidů * MeSH
- nádory tračníku enzymologie genetika metabolismus MeSH
- regulace genové exprese u nádorů * MeSH
- senioři MeSH
- stearyl-CoA-desaturasa genetika metabolismus MeSH
- syntázy mastných kyselin genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Flax (Linum usitatissimum L.) is grown for fiber and seed in many countries. Flax cultivars differ in the oil composition and, depending on the ratio of fatty acids, are used in pharmaceutical, food, or paint industries. It is known that genes of SAD (stearoyl-ACP desaturase) and FAD (fatty acid desaturase) families play a key role in the synthesis of fatty acids, and some alleles of these genes are associated with a certain composition of flax oil. However, data on genetic polymorphism of these genes are still insufficient. RESULTS: On the basis of the collection of the Institute for Flax (Torzhok, Russia), we formed a representative set of 84 cultivars and lines reflecting the diversity of fatty acid composition of flax oil. An approach for the determination of full-length sequences of SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes using the Illumina platform was developed and deep sequencing of the 6 genes in 84 flax samples was performed on MiSeq. The obtained high coverage (about 400x on average) enabled accurate assessment of polymorphisms in SAD1, SAD2, FAD2A, FAD2B, FAD3A, and FAD3B genes and evaluation of cultivar/line heterogeneity. The highest level of genetic diversity was observed for FAD3A and FAD3B genes - 91 and 62 polymorphisms respectively. Correlation analysis revealed associations between particular variants in SAD and FAD genes and predominantly those fatty acids whose conversion they catalyze: SAD - stearic and oleic acids, FAD2 - oleic and linoleic acids, FAD3 - linoleic and linolenic acids. All except one low-linolenic flax cultivars/lines contained both the substitution of tryptophan to stop codon in the FAD3A gene and histidine to tyrosine substitution in the FAD3B gene, while samples with only one of these polymorphisms had medium content of linolenic acid and cultivars/lines without them were high-linolenic. CONCLUSIONS: Genetic polymorphism of SAD and FAD genes was evaluated in the collection of flax cultivars and lines with diverse oil composition, and associations between particular polymorphisms and the ratio of fatty acids were revealed. The achieved results are the basis for the development of marker-assisted selection and DNA-based certification of flax cultivars.
- MeSH
- desaturasy mastných kyselin genetika MeSH
- DNA rostlinná MeSH
- genetická heterogenita MeSH
- genetická variace * MeSH
- kyselina alfa-linolenová metabolismus MeSH
- len enzymologie genetika metabolismus MeSH
- mastné kyseliny metabolismus MeSH
- oxygenasy se smíšenou funkcí genetika metabolismus MeSH
- rostlinné geny MeSH
- sekvenční analýza DNA MeSH
- substituce aminokyselin MeSH
- Publikační typ
- časopisecké články MeSH
Fatty acids are essential components of biological membranes, important for the maintenance of cellular structures, especially in organisms with complex life cycles like protozoan parasites. Apicomplexans are obligate parasites responsible for various deadly diseases of humans and livestock. We analyzed the fatty acids produced by the closest phototrophic relatives of parasitic apicomplexans, the chromerids Chromera velia and Vitrella brassicaformis, and investigated the genes coding for enzymes involved in fatty acids biosynthesis in chromerids, in comparison to their parasitic relatives. Based on evidence from genomic and metabolomic data, we propose a model of fatty acid synthesis in chromerids: the plastid-localized FAS-II pathway is responsible for the de novo synthesis of fatty acids reaching the maximum length of 18 carbon units. Short saturated fatty acids (C14:0-C18:0) originate from the plastid are then elongated and desaturated in the cytosol and the endoplasmic reticulum. We identified giant FAS I-like multi-modular enzymes in both chromerids, which seem to be involved in polyketide synthesis and fatty acid elongation. This full-scale description of the biosynthesis of fatty acids and their derivatives provides important insights into the reductive evolutionary transition of a phototropic algal ancestor to obligate parasites.
- MeSH
- Apicomplexa klasifikace genetika metabolismus MeSH
- biosyntetické dráhy genetika MeSH
- desaturasy mastných kyselin klasifikace genetika metabolismus MeSH
- druhová specificita MeSH
- elongasy mastných kyselin klasifikace genetika metabolismus MeSH
- fylogeneze MeSH
- lidé MeSH
- mastné kyseliny biosyntéza MeSH
- molekulární evoluce MeSH
- protozoální infekce parazitologie MeSH
- protozoální proteiny klasifikace genetika metabolismus MeSH
- synthasa mastných kyselin, typ 2 klasifikace genetika metabolismus MeSH
- synthasa mastných kyselin, typ I klasifikace genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Single nucleotide polymorphisms (SNPs) in FADS1/FADS2 genes are associated with changes in serum and tissue polyunsaturated fatty acid (PUFA) content. PUFA regulate inflammatory signaling pathways in adipose tissue; however, the effect of SNPs in FADS1/FADS2 on adipose tissue inflammation is equivocal. The present study examined if SNPs in FADS1/FADS2 modify human subcutaneous adipose tissue (SAT) fatty acid profiles and the expression of genes associated with inflammation/immune function, lipid metabolism, and cellular differentiation. METHODS: SAT fatty acids and the expression of 117 genes were measured in 174 men and women from the DiOGenes Study using gas chromatography and qRT-PCR, respectively. Associations between fatty acids, gene expression, and SNPs in FADS1/FADS2 were investigated by linear regression and multivariate analysis. RESULTS: Four SNPs (rs174537, rs174546, rs174556, rs174601) in FADS1/FADS2 were significantly associated with SAT fatty acids. All SNPs were in high linkage disequilibrium with the commonly reported rs174537 SNP in FADS1. Minor allele carriers for rs174537 (GT+TT) had reduced 20:4n-6 (p = 1.74E-5), lower delta-5 desaturase enzyme activity (p = 2.09E-9), and lower FADS1 gene expression (p = 0.03) compared to major GG carriers. Multivariate analysis revealed that 20:4n-6 and 20:3n-6 explained ~19% of the variance between rs174537 genotypes, while gene expression explained <7%. Receiver operating characteristic (ROC) curves indicated that rs174537 genotype can be distinguished with SAT fatty acids (AUC = 0.842), but not gene expression (AUC = 0.627). No differences in SAT inflammatory gene expression were observed between rs174537 genotypes. SAT 20:3n-6 levels were positively correlated with the expression of several inflammatory genes, and inversely correlated with FADS1 expression. CONCLUSION: This study showed that FADS1 genotype is distinguished by SAT fatty acid profiles, but not inflammatory gene expression.
- MeSH
- buněčná diferenciace genetika MeSH
- desaturasy mastných kyselin genetika metabolismus MeSH
- dospělí MeSH
- exprese genu MeSH
- genotyp MeSH
- imunitní systém MeSH
- jednonukleotidový polymorfismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- lineární modely MeSH
- mastné kyseliny genetika MeSH
- metabolismus lipidů genetika MeSH
- multigenová rodina genetika MeSH
- multivariační analýza MeSH
- obezita genetika MeSH
- podkožní tuk metabolismus MeSH
- zánět genetika MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
After cold shock, the Bacillus subtilis desaturase Des introduces double bonds into the fatty acids of existing membrane phospholipids. The synthesis of Des is regulated exclusively by the two-component system DesK/DesR; DesK serves as a sensor of the state of the membrane and triggers Des synthesis after a decrease in membrane fluidity. The aim of our work is to investigate the biophysical changes in the membrane that are able to affect the DesK signalling state. Using linear alcohols (ethanol, propanol, butanol, hexanol, octanol) and benzyl alcohol, we were able to suppress Des synthesis after a temperature downshift. The changes in the biophysical properties of the membrane caused by alcohol addition were followed using membrane fluorescent probes and differential scanning calorimetry. We found that the membrane fluidization induced by alcohols was reflected in an increased hydration at the lipid-water interface. This is associated with a decrease in DesK activity. The addition of alcohol mimics a temperature increase, which can be measured isothermically by fluorescence anisotropy. The effect of alcohols on the membrane periphery is in line with the concept of the mechanism by which two hydrophilic motifs located at opposite ends of the transmembrane region of DesK, which work as a molecular caliper, sense temperature-dependent variations in membrane properties.
- MeSH
- alkoholy farmakologie MeSH
- aminokyselinové motivy MeSH
- Bacillus subtilis metabolismus MeSH
- bakteriální proteiny metabolismus MeSH
- buněčná membrána účinky léků fyziologie MeSH
- desaturasy mastných kyselin biosyntéza genetika MeSH
- diferenciální skenovací kalorimetrie MeSH
- enzymová indukce účinky léků MeSH
- fluidita membrány účinky léků MeSH
- fluorescenční polarizace MeSH
- fosforylace MeSH
- hydrofobní a hydrofilní interakce MeSH
- mastné kyseliny metabolismus MeSH
- nízká teplota MeSH
- posttranslační úpravy proteinů * MeSH
- proteinkinasy metabolismus MeSH
- regulace genové exprese u bakterií účinky léků MeSH
- rekombinantní fúzní proteiny metabolismus MeSH
- reportérové geny MeSH
- signální transdukce účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Stavy asociované s inzulinovou rezistencí, jako jsou nadváha/obezita, diabetes mellitus 2. typu (DM2), kardiovaskulární onemocnění (KVO), některé nádory a neurologicko-psychiatrické choroby, jsou charakterizovány poklesem obsahu vícenenasycených mastných kyselin s dlouhým řetězcem (LC-PUFA). Množství LC-PUFA závisí jednak na exogenním příjmu jejich prekurzorů – kyseliny linolové a α-linolenové – a dále na rychlosti jejich metabolické přeměny, na níž se podílí řada enzymů: delta-6-desaturáza (D6D, FADS2), D5D (FADS1) a elongázy (Elovl2, -5, 6). Změny aktivit D5D/D6D byly popsány u řady chorob, mezi něž patří neurologicko-psychiatrická onemocnění (depresivní poruchy, bipolární afektivní poruchy, demence), metabolická onemocnění (obezita, metabolický syndrom, DM2), KVO (hypertenze, ischemická choroba srdeční), chronické záněty a alergie (Crohnova nemoc, atopický ekzém) a některé nádory. Analogické výsledky byly prokázány asociačními studiemi mezi genotypy/haplotypy FADS1/FADS2 a výše uvedenými skupinami onemocnění, případně interakce mezi příjmem kyseliny linolové (LA) i α-linolenové (ALA) a výskytem minoritních alel polymorfismů FADS1/FADS2, které mají vesměs nižší enzymatické aktivity. Pokles aktivit desaturáz se projeví sníženou koncentrací produktů spojenou se zvýšenou koncentrací substrátu. Byly popsány asociace některých SNPFADSs ischemickou chorobou srdeční, koncentracemi lipidů, oxidačním stresem, glukózovou homeostázou i zánětlivou reakcí. Experimentální studie na zvířecích modelech a výskyt vzácných onemocnění spojených s chyběním, resp. výrazným poklesem aktivit D5D/D6D vyzdvihly význam desaturáz pro zdravý vývoj organismu i pro patogenezi některých chorob. Klíčová slova: delta-5-desaturáza, delta-6-desaturáza, geny FADS1/FADS2, vícenenasycené mastné kyseliny, zánět, oxidační stres, kardiovaskulární a metabolické choroby
States associated with insulin resistance, as overweight/obesity, type 2 diabetes mellitus (DM2), cardiovascular diseases (CVD), some cancers and neuropsychiatric diseases are characterized with a decrease of long-chain polyunsaturated fatty acids (LC-PUFA) levels. Amounts of LC-PUFA depend on the exogenous intake of their precursors [linoleic (LA) and α-linolenic acid (ALA)] and by rate of their metabolism, which is influenced by activities of enzymes, such as Δ6-desaturase (D6D, FADS2), D5D, FADS1, elongases (Elovl2, -5, 6). Altered activities of D5D/D6D were described in plenty of diseases, e.g. neuropsychiatric (depressive disorders, bipolar disorder, dementia), metabolic (obesity, metabolic syndrome, DM2) and cardiovascular diseases (arterial hypertension, coronary heart disease), inflammatory states and allergy (Crohn’s disease, atopic eczema) or some malignancies. Similar results were obtained in studies dealing with the associations between genotypes/haplotypes of FADS1/FADS2 and above mentioned diseases, or interactions of dietary intake of LA and ALA on one hand and of the polymorphisms of minor allels of FADS1/FADS2, usually characterized by lower activities, on the other hand. The decrease of the desaturases activities leads to decreased concentrations of products with concomitant increased concentrations of substrates. Associations of some SNPFADSwith coronary heart disease, concentrations of plasma lipids, oxidative stress, glucose homeostasis, and inflammatory reaction, were described. Experimental studies on animal models and occurrence of rare diseases, associated with missing or with marked fall activities of D5D/D6D emphasized the significance of desaturases for healthy development of organism as well as for pathogenesis of some disease. Keywords: delta-5-desaturase, delta-6-desaturase, genes FADS1/FADS2, polyunsaturated fatty acids, inflammation, oxidative stress, cardiovascular and metabolic diseases
- Klíčová slova
- delta-5-desaturáza, vícenenasycené mastné kyseliny, geny FADS1/FADS2, delta-6-desaturáza,
- MeSH
- desaturasy mastných kyselin * genetika MeSH
- diabetes mellitus 2. typu genetika MeSH
- dyslipidemie genetika MeSH
- inzulinová rezistence genetika MeSH
- ischemická choroba srdeční genetika MeSH
- kardiovaskulární nemoci genetika MeSH
- kyselina linolová metabolismus MeSH
- lidé MeSH
- metabolický syndrom genetika MeSH
- multigenová rodina MeSH
- nenasycené mastné kyseliny * fyziologie genetika metabolismus MeSH
- oxidační stres MeSH
- polymorfismus genetický * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
For sexual communication, moths primarily use blends of fatty acid derivatives containing one or more double bonds in various positions and configurations, called sex pheromones (SPs). To study the molecular basis of novel SP component (SPC) acquisition, we used the tobacco hornworm (Manduca sexta), which uses a blend of mono-, di-, and uncommon triunsaturated fatty acid (3UFA) derivatives as SP. We identified pheromone-biosynthetic fatty acid desaturases (FADs) MsexD3, MsexD5, and MsexD6 abundantly expressed in the M. sexta female pheromone gland. Their functional characterization and in vivo application of FAD substrates indicated that MsexD3 and MsexD5 biosynthesize 3UFAs via E/Z14 desaturation from diunsaturated fatty acids produced by previously characterized Z11-desaturase/conjugase MsexD2. Site-directed mutagenesis of sequentially highly similar MsexD3 and MsexD2 demonstrated that swapping of a single amino acid in the fatty acyl substrate binding tunnel introduces E/Z14-desaturase specificity to mutated MsexD2. Reconstruction of FAD gene phylogeny indicates that MsexD3 was recruited for biosynthesis of 3UFA SPCs in M. sexta lineage via gene duplication and neofunctionalization, whereas MsexD5 representing an alternative 3UFA-producing FAD has been acquired via activation of a presumably inactive ancestral MsexD5. Our results demonstrate that a change as small as a single amino acid substitution in a FAD enzyme might result in the acquisition of new SP compounds.
- MeSH
- desaturasy mastných kyselin genetika metabolismus MeSH
- fylogeneze MeSH
- hmyzí proteiny genetika metabolismus MeSH
- Manduca genetika metabolismus MeSH
- molekulární evoluce * MeSH
- molekulární sekvence - údaje MeSH
- nenasycené mastné kyseliny genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sexuální lákadla biosyntéza genetika MeSH
- substituce aminokyselin * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- desaturasy mastných kyselin * farmakologie genetika metabolismus MeSH
- inzulinová rezistence * genetika MeSH
- ischemická choroba srdeční dietoterapie metabolismus prevence a kontrola MeSH
- lidé MeSH
- metabolický syndrom dietoterapie metabolismus prevence a kontrola MeSH
- rizikové faktory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- desaturasy mastných kyselin genetika metabolismus MeSH
- lidé MeSH
- mastné kyseliny metabolismus MeSH
- metabolický syndrom * imunologie metabolismus MeSH
- nenasycené mastné kyseliny terapeutické užití MeSH
- tuková tkáň enzymologie imunologie MeSH
- zánět imunologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
