Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 702
Filtrar
1.
PLoS One ; 15(2): e0226888, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32027663

RESUMO

Camellia oleifera Abel. (C. oleifera), as an important woody tree species producing edible oils in China, has attracted enormous attention due to its abundant unsaturated fatty acids and their associated benefits to human health. To reveal novel insights into the characters during the maturation period of this plant as well as the molecular basis of fatty acid biosynthesis and degradation, we conducted a conjoint analysis of the transcriptome and proteome of C. oleifera seeds from Hainan Island. Using RNA sequencing (RNA-seq) technology and shotgun proteomic method, 59,391 transcripts and 40,500 unigenes were obtained by TIGR Gene Indices Clustering Tools (TGICL), while 1691 protein species were identified from Mass Spectrometry (MS). Subsequently, all genes and proteins were employed in euKaryotic Orthologous Groups (KOG) classification, Gene Ontology (GO) annotation, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to investigate their essential functions. The results indicated that the most abundant pathways were biological metabolic processes. There were 946 unigenes associated with lipid metabolism at the transcriptome level, with 116 proteins at the proteome level; among these, 38 specific proteins were involved in protein-protein interactions, with the majority being related to fatty acid catabolic process. The expression levels of 21 candidate unigenes encoding target proteins were further detected by quantitative real-time polymerase chain reaction (qRT-PCR). Finally, Gas Chromatography Mass Spectrometry (GC-MS) was carried out to determine the fatty acid composition of C. oleifera oil. These findings not only deepened our understanding about the molecular mechanisms of fatty acid metabolism but also offered new evidence concerning the roles of relevant proteins in oil-bearing crops. Furthermore, the lipid-associated proteins recognized in this research might be helpful in providing a reference for the synthetic regulation of C. oleifera oil quality by genetic engineering techniques, thus resulting in potential application in agriculture.


Assuntos
Camellia/genética , Ácidos Graxos/genética , Metabolismo dos Lipídeos/genética , Proteoma/metabolismo , Sementes/genética , Transcriptoma/genética , China , Perfilação da Expressão Gênica/métodos , Ilhas , Óleos Vegetais/química , Análise de Sequência de RNA/métodos
2.
Nat Commun ; 11(1): 45, 2020 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-31896749

RESUMO

Unlike protein-coding genes, the majority of human long non-coding RNAs (lncRNAs) are considered non-conserved. Although lncRNAs have been shown to function in diverse pathophysiological processes in mice, it remains largely unknown whether human lncRNAs have such in vivo functions. Here, we describe an integrated pipeline to define the in vivo function of non-conserved human lncRNAs. We first identify lncRNAs with high function potential using multiple indicators derived from human genetic data related to cardiometabolic traits, then define lncRNA's function and specific target genes by integrating its correlated biological pathways in humans and co-regulated genes in a humanized mouse model. Finally, we demonstrate that the in vivo function of human-specific lncRNAs can be successfully examined in the humanized mouse model, and experimentally validate the predicted function of an obesity-associated lncRNA, LINC01018, in regulating the expression of genes in fatty acid oxidation in humanized livers through its interaction with RNA-binding protein HuR.


Assuntos
Fígado/fisiologia , RNA Longo não Codificante/fisiologia , Animais , Sequência de Bases , Sequência Conservada , Proteína Semelhante a ELAV 1/genética , Proteína Semelhante a ELAV 1/metabolismo , Epigênese Genética , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Estudo de Associação Genômica Ampla , Hepatócitos/fisiologia , Humanos , Fígado/metabolismo , Hepatopatias/genética , Hepatopatias/metabolismo , Masculino , Metiltransferases/genética , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Obesidade/genética , Obesidade/metabolismo , Locos de Características Quantitativas
3.
Cell Prolif ; 53(1): e12705, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31657086

RESUMO

OBJECTIVES: Increasing evidences demonstrate a close correlation between epithelial-to-mesenchymal transition (EMT) induction and cancer lipid metabolism. However, the molecular mechanisms have not been clarified. MATERIALS AND METHODS: In our study, the relative expression level of PRRX1 was detected, its relationship with free fatty acid (FFA) and PPARG2 was analysed in 85 SACC tissues and 15 salivary glands from the benign salivary tumours. We also compared the FFAs composition and levels in these SACC cells. PPARG2 was detected in PRRX1-induced FFAs treatment as well as Src and MMP-9 were detected in FFAs treatment-induced invasion and migration of SACC cells, and ChIP test was performed to identify the target interactions. RESULTS: Our data showed that overexpression of PRRX1 induced EMT and facilitated the invasion and migration of SACC cells, and PRRX1 expression was closely associated with high FFAs level and poor prognosis of SACC patients. Furthermore, PRRX1 silence led to the increase of PPARG2 and the reduction of FFAs level and the migration and invasion of SACC cells. And inhibition of PPARG2 rescued FFAs level and migration and invasion capabilities of SACC cells. Free fatty acids treatment induced an increase of Stat5-DNA binding activity via Src- and MMP-9-dependent pathway. CONCLUSIONS: Collectively, our findings showed that the PRRX1/PPARG2/FFAs signalling in SACC was important for accelerating tumour metastasis through the induction of EMT and the metabolic reprogramming of FFAs.


Assuntos
Carcinoma Adenoide Cístico/metabolismo , Reprogramação Celular , Transição Epitelial-Mesenquimal , Ácidos Graxos/metabolismo , Proteínas de Homeodomínio/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias das Glândulas Salivares/metabolismo , Animais , Carcinoma Adenoide Cístico/genética , Carcinoma Adenoide Cístico/patologia , Linhagem Celular Tumoral , Ácidos Graxos/genética , Proteínas de Homeodomínio/genética , Humanos , Camundongos , Invasividade Neoplásica , Metástase Neoplásica , Proteínas de Neoplasias/genética , Neoplasias das Glândulas Salivares/genética , Neoplasias das Glândulas Salivares/patologia
4.
PLoS Genet ; 15(11): e1008375, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31738765

RESUMO

DNA variants that alter gene expression contribute to variation in many phenotypic traits. In particular, trans-acting variants, which are often located on different chromosomes from the genes they affect, are an important source of heritable gene expression variation. However, our knowledge about the identity and mechanism of causal trans-acting variants remains limited. Here, we developed a fine-mapping strategy called CRISPR-Swap and dissected three expression quantitative trait locus (eQTL) hotspots known to alter the expression of numerous genes in trans in the yeast Saccharomyces cerevisiae. Causal variants were identified by engineering recombinant alleles and quantifying the effects of these alleles on the expression of a green fluorescent protein-tagged gene affected by the given locus in trans. We validated the effect of each variant on the expression of multiple genes by RNA-sequencing. The three variants differed in their molecular mechanism, the type of genes they reside in, and their distribution in natural populations. While a missense leucine-to-serine variant at position 63 in the transcription factor Oaf1 (L63S) was almost exclusively present in the reference laboratory strain, the two other variants were frequent among S. cerevisiae isolates. A causal missense variant in the glucose receptor Rgt2 (V539I) occurred at a poorly conserved amino acid residue and its effect was strongly dependent on the concentration of glucose in the culture medium. A noncoding variant in the conserved fatty acid regulated (FAR) element of the OLE1 promoter influenced the expression of the fatty acid desaturase Ole1 in cis and, by modulating the level of this essential enzyme, other genes in trans. The OAF1 and OLE1 variants showed a non-additive genetic interaction, and affected cellular lipid metabolism. These results demonstrate that the molecular basis of trans-regulatory variation is diverse, highlighting the challenges in predicting which natural genetic variants affect gene expression.


Assuntos
Proteínas de Ligação a DNA/genética , Evolução Molecular , Sequências Reguladoras de Ácido Nucleico/genética , Proteínas de Saccharomyces cerevisiae/genética , Estearoil-CoA Dessaturase/genética , Fatores de Transcrição/genética , Sistemas CRISPR-Cas/genética , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Regulação Fúngica da Expressão Gênica/genética , Proteínas de Fluorescência Verde/genética , Metabolismo dos Lipídeos/genética , Proteínas de Transporte de Monossacarídeos/genética , Mutação de Sentido Incorreto/genética , Fenótipo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo
5.
Nat Commun ; 10(1): 4788, 2019 10 21.
Artigo em Inglês | MEDLINE | ID: mdl-31636271

RESUMO

Genetic studies of metabolites have identified thousands of variants, many of which are associated with downstream metabolic and obesogenic disorders. However, these studies have relied on univariate analyses, reducing power and limiting context-specific understanding. Here we aim to provide an integrated perspective of the genetic basis of metabolites by leveraging the Finnish Metabolic Syndrome In Men (METSIM) cohort, a unique genetic resource which contains metabolic measurements, mostly lipids, across distinct time points as well as information on statin usage. We increase effective sample size by an average of two-fold by applying the Covariates for Multi-phenotype Studies (CMS) approach, identifying 588 significant SNP-metabolite associations, including 228 new associations. Our analysis pinpoints a small number of master metabolic regulator genes, balancing the relative proportion of dozens of metabolite levels. We further identify associations to changes in metabolic levels across time as well as genetic interactions with statin at both the master metabolic regulator and genome-wide level.


Assuntos
Pleiotropia Genética , Síndrome Metabólica/genética , Metaboloma/genética , Idoso , Aminoácidos/genética , Aminoácidos/metabolismo , Estudos de Coortes , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Redes Reguladoras de Genes , Estudo de Associação Genômica Ampla , Humanos , Lipoproteínas HDL/genética , Lipoproteínas HDL/metabolismo , Lipoproteínas IDL/genética , Lipoproteínas IDL/metabolismo , Lipoproteínas LDL/genética , Lipoproteínas LDL/metabolismo , Lipoproteínas VLDL/genética , Lipoproteínas VLDL/metabolismo , Espectroscopia de Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único
6.
Mar Drugs ; 17(9)2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31484443

RESUMO

Phytoplankton are primary producers in the marine ecosystem, where phosphorus is often a limiting factor of their growth. Hence, they have evolved strategies to recycle phosphorus by replacing membrane phospholipids with phosphorus-free lipids. However, mechanisms for replacement of lipid classes remain poorly understood. To improve our understanding, we performed the lipidomic and transcriptomic profiling analyses of an oleaginous marine microalga Nannochloropsis sp. PJ12 in response to phosphorus depletion (PD) and replenishing. In this study, by using (liquid chromatography couple with tandem mass spectrometry) LC-MS/MS-based lipidomic analysis, we show that membrane phospholipid levels are significantly reduced upon PD, while phosphorus-free betaine lipid levels are increased. However, levels of phosphorus-free photosynthetic galactolipid and sulfolipid are not increased upon PD, consistent with the reduced photosynthetic activity. RNA-seq-based transcriptomic analysis indicates that enzymes involved in phospholipid recycling and phosphorus-free lipid synthesis are upregulated, supporting the lipidomic analysis. Furthermore, enzymes involved in FASII (type II fatty acid synthesis) elongation cycle upon PD are transcriptionally downregulated. EPA (eicosapentaenoic acid) level decrease upon PD is revealed by both GC-MS (gas chromatography coupled with mass spectrometry) and LC-MS/MS-based lipidomic analyses. PD-induced alteration is reversed after phosphorus replenishing. Taken together, our results suggest that the alteration of lipid classes upon environmental change of phosphorus is a result of remodeling rather than de novo synthesis in Nannochloropsis sp. PJ12.


Assuntos
Metabolismo dos Lipídeos/efeitos dos fármacos , Microalgas/efeitos dos fármacos , Fósforo/farmacologia , Transcriptoma/efeitos dos fármacos , Cromatografia Líquida/métodos , Ácidos Graxos/genética , Perfilação da Expressão Gênica/métodos , Glicolipídeos/genética , Metabolismo dos Lipídeos/genética , Lipídeos/genética , Microalgas/genética , Fosfolipídeos/genética , Fotossíntese/efeitos dos fármacos , Fotossíntese/genética , Fitoplâncton/efeitos dos fármacos , Fitoplâncton/genética , Espectrometria de Massas em Tandem/métodos , Transcriptoma/genética
7.
J Biosci ; 44(4)2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31502567

RESUMO

Constantly rising energy demands, finite fossil fuel reserves and deteriorating environmental conditions have invoked worldwide interest to explore the sustainable sources of renewable biofuels. Locally adapted photosynthetic oleaginous microalgae with rapid growth on variable temperatures could be an ideal way for bioremediating the wastewater (WW) while producing the feedstock for biodiesel. To test this notion, an unknown strain was isolated from a sewage fed lake (Neela-Hauz). It was discerned as Chlorella sorokiniana-I using the 16S rDNA and 18S rDNA barcodes. The culture conditions such as pH, illumination, different temperature ranges and growth medium were cohesively optimized prior to the assessment of C. sorokiniana-I's efficacy to remediate the WWand biodiesel production. The strain has thrived well up to 40°C when continuously grown for 15 days. The highest lipid accumulation and biomass productivity were recorded in 100% WW. Fatty acid methyl ester (FAME) content was observed to be more than twice in WW (47%), compared to control synthetic media, TAP (20%) and BG11 (10%), which indicate the importance of this new isolate for producing economically viable biodiesel. Moreover, it is highly efficient in removing the total nitrogen (77%), total phosphorous (81%), iron (67%) and calcium (42%) from the WW. The quality of WW was considerably improved by reducing the overall chemical oxygen demand (48%), biological oxygen demand (47%) and alkalinity (15%). Thus, C. sorokiniana-I could be an ideal alga for the tropical countries in the remediation of WW while producing feedstock for biodiesel in a cost-effective manner.


Assuntos
Biodegradação Ambiental , Chlorella/genética , Recuperação e Remediação Ambiental , Estágios do Ciclo de Vida/genética , Biocombustíveis , Biomassa , Chlorella/crescimento & desenvolvimento , Chlorella/metabolismo , Meios de Cultura , Ácidos Graxos/química , Ácidos Graxos/genética , Humanos , Lipídeos/química , Lipídeos/genética , Microalgas , Nitrogênio/metabolismo , Águas Residuárias/química
8.
Microb Cell Fact ; 18(1): 135, 2019 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-31409350

RESUMO

BACKGROUND: Medium-chain-length polyhydroxyalkanoates (mcl-PHAs) containing various chain length monomers from C6 to C14 have more applications besides sustainable and environmental-friendly biomaterials owing to their superior physical and mechanical properties. We engineered a reversed fatty acid ß-oxidation pathway in Escherichia coli that can synthesize mcl-PHA directly from glucose and achieved high yield. However, there were only even-chain monomers in the biosynthetic polymers. The need for mcl-PHA harboring both even- and odd-chain monomers with better and wider utility impels us to develop the biosynthetic routes for the production of the novel and unnatural mcl-PHA through rewiring the basic metabolism. RESULTS: In the present study, a propionate assimilation and metabolic route was integrated into the reversed fatty acid ß-oxidation in order to produce mcl-PHA consisting of both even- and odd-numbered monomers. The content of odd-numbered monomers in mcl-PHA was improved with the increased propionate addition. After further deletion of pyruvate oxidase (PoxB) and pyruvate formate-lyase (PflB), the metabolically engineered chassis E. coli LZ08 harboring pQQ05 and pZQ06 (overexpression of prpP and prpE genes from Ralstonia eutropha H16) innovatively accumulated 6.23 wt% mcl-PHA containing odd-chain monomers ranging from 7 to 13 carbon atoms about 20.03 mol%. CONCLUSIONS: This is the first successful report on production of mcl-PHA harboring both even- and odd-chain monomers (C6-C14) synthesized from glucose and propionate in recombinant E. coli. This present study achieved the highest yield of de novo production of mcl-PHA containing odd-numbered monomers in E. coli at shake-flask fermentation level. Continued engineering of host strains and pathway enzymes will ultimately lead to more economical production of odd-chain monomers based on market demand. The synthetic pathway can provide a promising platform for production of other value-added chemicals and biomaterials that use acetyl-CoA and propionyl-CoA as versatile precursors and can be extended to other microorganisms as intelligent cell factories.


Assuntos
Escherichia coli/genética , Escherichia coli/metabolismo , Engenharia Metabólica , Poli-Hidroxialcanoatos/biossíntese , Vias Biossintéticas , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Regulação Bacteriana da Expressão Gênica , Glucose/metabolismo , Oxirredução
9.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1864(12): 158513, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31465888

RESUMO

The biosynthetic pathways for most lipophilic metabolites share several common principles. These substances are built almost exclusively from acetyl-CoA as the donor for the carbon scaffold and NADPH is required for the reductive steps during biosynthesis. Due to their hydrophobicity, the end products are sequestered into the same cellular compartment, the lipid droplet. In this review, we will summarize the efforts in the metabolic engineering of yeasts for the production of two major hydrophobic substance classes, fatty acid-based lipids and isoprenoids, with regard to these common aspects. We will compare and discuss the results of genetic engineering strategies to construct strains with enhanced synthesis of the precursor acetyl-CoA and with modified redox metabolism for improved NADPH supply. We will also discuss the role of the lipid droplet in the storage of the hydrophobic product and review the strategies to either optimize this organelle for higher capacity or to achieve excretion of the product into the medium.


Assuntos
Ácidos Graxos/genética , Hemiterpenos/genética , Engenharia Metabólica/métodos , Leveduras/genética , Acetilcoenzima A/genética , Acetilcoenzima A/metabolismo , Vias Biossintéticas , Butadienos/metabolismo , Ácidos Graxos/metabolismo , Hemiterpenos/metabolismo , Microbiologia Industrial/métodos , Metabolismo dos Lipídeos , NADP/genética , NADP/metabolismo , Leveduras/metabolismo
10.
PLoS One ; 14(7): e0218895, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31329601

RESUMO

The tenacious human parasitic helminth Strongyloides stercoralis is a significant health problem worldwide. The current lack of a definitive diagnostic laboratory test to rule out this infection necessitates designing more specific diagnostic methods. Fatty acid and retinol-binding protein (FAR) plays a crucial role in the development and reproduction of nematodes. We generated a recombinant form of this protein and determined its applicability for immunodiagnosis of S. stercoralis. The L3 form of S. stercoralis was harvested and used for RNA extraction and cDNA synthesis. The coding sequence of S. stercoralis FAR (SsFAR) was cloned into pET28a(+) vector, expressed in E. coli BL21 and purified. ELISA and immunoblotting were employed to determine the specificity and sensitivity of rSsFAR using a set of defined sera. In addition, we analyzed the phylogenetic relationship of SsFAR with different FAR sequences from other nematodes. The cloned SsFAR had an open reading frame of 447 bp encoding 147 amino acids, with a deduced molecular mass of 19 kD. The SsFAR amino acid sequence was 93% identical to FAR of S. ratti. For differential immunodiagnosis of strongyloidiasis, rSsFAR exhibited 100% sensitivity and 97% specificity. However, cross-reactivity with FAR proteins of other parasites, namely Toxocara canis and Echinococcus granulosus, was noted. Our results provide a novel approach for immunodiagnosis of S. stercoralis infections using rSsFAR with reliable sensitivity and specificity.


Assuntos
Proteínas Recombinantes/genética , Proteínas de Ligação ao Retinol/genética , Strongyloides stercoralis/genética , Estrongiloidíase/diagnóstico , Animais , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Antígenos de Helmintos/isolamento & purificação , Testes Diagnósticos de Rotina , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Humanos , Testes Imunológicos/métodos , Filogenia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Proteínas de Ligação ao Retinol/isolamento & purificação , Strongyloides stercoralis/imunologia , Strongyloides stercoralis/patogenicidade , Estrongiloidíase/genética , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologia
11.
G3 (Bethesda) ; 9(9): 2963-2975, 2019 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-31296616

RESUMO

Oat (Avena sativa L.) has a high concentration of oils, comprised primarily of healthful unsaturated oleic and linoleic fatty acids. To accelerate oat plant breeding efforts, we sought to identify loci associated with variation in fatty acid composition, defined as the types and quantities of fatty acids. We genotyped a panel of 500 oat cultivars with genotyping-by-sequencing and measured the concentrations of ten fatty acids in these oat cultivars grown in two environments. Measurements of individual fatty acids were highly correlated across samples, consistent with fatty acids participating in shared biosynthetic pathways. We leveraged these phenotypic correlations in two multivariate genome-wide association study (GWAS) approaches. In the first analysis, we fitted a multivariate linear mixed model for all ten fatty acids simultaneously while accounting for population structure and relatedness among cultivars. In the second, we performed a univariate association test for each principal component (PC) derived from a singular value decomposition of the phenotypic data matrix. To aid interpretation of results from the multivariate analyses, we also conducted univariate association tests for each trait. The multivariate mixed model approach yielded 148 genome-wide significant single-nucleotide polymorphisms (SNPs) at a 10% false-discovery rate, compared to 129 and 73 significant SNPs in the PC and univariate analyses, respectively. Thus, explicit modeling of the correlation structure between fatty acids in a multivariate framework enabled identification of loci associated with variation in seed fatty acid concentration that were not detected in the univariate analyses. Ultimately, a detailed characterization of the loci underlying fatty acid variation can be used to enhance the nutritional profile of oats through breeding.


Assuntos
Avena/genética , Ácidos Graxos/genética , Estudo de Associação Genômica Ampla/métodos , Sementes/genética , Sementes/metabolismo , Avena/metabolismo , Ácidos Graxos/metabolismo , Genética Populacional , Genoma de Planta , Fenótipo , Polimorfismo de Nucleotídeo Único
12.
PLoS One ; 14(7): e0219435, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31291335

RESUMO

Carrier proteins are four-helix bundles that covalently hold metabolites and secondary metabolites, such as fatty acids, polyketides and non-ribosomal peptides. These proteins mediate the production of many pharmaceutically important compounds including antibiotics and anticancer agents. Acyl carrier proteins (ACPs) can be found as part of a multi-domain polypeptide (Type I ACPs), or as part of a multiprotein complex (Type II). Here, the main focus is on ACP2 and ACP3, domains from the type I trans-AT polyketide synthase MmpA, which is a core component of the biosynthetic pathway of the antibiotic mupirocin. During molecular dynamics simulations of their apo, holo and acyl forms ACP2 and ACP3 both form a substrate-binding surface-groove. The substrates bound to this surface-groove have polar groups on their acyl chain exposed and forming hydrogen bonds with the solvent. Bulky hydrophobic residues in the GXDS motif common to all ACPs, and similar residues on helix III, appear to prohibit the formation of a deep tunnel in type I ACPs and type II ACPs from polyketide synthases. In contrast, the equivalent positions in ACPs from type II fatty acid synthases, which do form a deep solvent-excluded substrate-binding tunnel, have the small residue alanine. During simulation, ACP3 with mutations I61A L36A W44L forms a deep tunnel that can fully bury a saturated substrate in the core of the ACP, in contrast to the surface groove of the wild type ACP3. Similarly, in the ACP from E. coli fatty acid synthase, a type II ACP, mutations can change ligand binding from being inside a deep tunnel to being in a surface groove, thus demonstrating how changing a few residues can modify the possibilities for ligand binding.


Assuntos
Proteína de Transporte de Acila/química , Complexos Multiproteicos/química , Peptídeos/química , Policetídeo Sintases/química , Acinetobacter baumannii/química , Acinetobacter baumannii/genética , Proteína de Transporte de Acila/genética , Proteína de Transporte de Acila/metabolismo , Motivos de Aminoácidos/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Vias Biossintéticas/genética , Sequestro de Carbono/genética , Escherichia coli/genética , Ácido Graxo Sintase Tipo II/química , Ácido Graxo Sintase Tipo II/genética , Ácido Graxo Sintase Tipo II/metabolismo , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Simulação de Dinâmica Molecular , Complexos Multiproteicos/genética , Mupirocina/biossíntese , Mupirocina/metabolismo , Peptídeos/genética , Mutação Puntual/genética , Policetídeo Sintases/genética , Ligação Proteica
13.
Mar Biotechnol (NY) ; 21(5): 643-654, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31273567

RESUMO

Asian seabass is an important food fish species. While improving growth, increasing the nutritional value is important, omega-3 fatty acids are indispensable to human health. Identifying and validating DNA markers associated with traits is the first step towards marker-assisted selection (MAS). We quantified 13 different fatty acids and three growth traits in 213 F2 Asian seabass from a family at the age 270 days post hatch, and screened QTL for these traits. The content of total fatty acids in 100 g flesh was 2.57 ± 0.80 g, while the proportions of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were 16.96 ± 2.20% and 5.42 ± 0.90%, respectively. A linkage map with 2424 SNPs was constructed and used for QTL mapping. For fatty acid compositions, 14 significant QTL were identified on three linkage groups (LG5, LG11 and LG14), with phenotypic variance explained (PVE) from 12.8 to 24.6%. Thirty-nine suggestive QTL were detected on 16 LGs. Two significant QTL for EPA were identified on LG5 and LG14, with PVE of 15.2% and 15.1%, respectively. No significant QTL was identified for DHA. For growth traits, six significant and 13 suggestive QTL were identified on two and seven LGs, respectively. Only a few significant QTL for fatty acids overlapped with previously mapped QTL for these traits, suggesting that most QTL detected in a family are family-specific and could only be used in MAS in the family per se. To facilitate population-wide molecular breeding, more powerful methods (e.g. GWAS) should be used to identify SNPs for genomic selection.


Assuntos
Bass/genética , Ácidos Docosa-Hexaenoicos/genética , Ácido Eicosapentaenoico/genética , Genoma , Locos de Características Quantitativas , Característica Quantitativa Herdável , Animais , Bass/crescimento & desenvolvimento , Bass/metabolismo , Mapeamento Cromossômico/métodos , Ácidos Docosa-Hexaenoicos/biossíntese , Ácido Eicosapentaenoico/biossíntese , Ácidos Graxos/biossíntese , Ácidos Graxos/classificação , Ácidos Graxos/genética , Ligação Genética , Genótipo , Músculos/metabolismo , Polimorfismo de Nucleotídeo Único
14.
Nutrients ; 11(7)2019 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-31331006

RESUMO

Available evidence on the associations of dietary and circulating levels of long-chain n-3 fatty acids, which have potential antiarrhythmic properties, and other fatty acids with atrial fibrillation is conflicting and limited. We conducted a Mendelian randomization study to assess the associations between plasma phospholipid fatty acid levels and atrial fibrillation. Summary-level data of atrial fibrillation were available from 65,446 cases and 522,744 non-cases included in the Atrial Fibrillation Consortium. Sixteen single-nucleotide polymorphisms associated with ten fatty acids at significance level of p < 5 × 10-8 were identified as instrumental variables from the hitherto largest genome-wide association studies for plasma fatty acids. The fixed-effects inverse-variance weighted method was used to assess the association of individual plasma fatty acids and atrial fibrillation risk. The random-effects inverse-variance weighted method, weighted median method, and Mendelian randomization (MR)-Egger method were employed as the sensitivity analyses. Genetic predisposition to higher levels of any of the ten individual fatty acids was not associated with atrial fibrillation risk.


Assuntos
Fibrilação Atrial/sangue , Ácidos Graxos/sangue , Análise da Randomização Mendeliana , Fosfolipídeos/sangue , Fibrilação Atrial/genética , Ácidos Graxos/genética , Ácidos Graxos Ômega-3/sangue , Ácidos Graxos Ômega-3/genética , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla , Humanos , Ácido Linoleico/sangue , Ácido Linoleico/genética , Polimorfismo de Nucleotídeo Único/genética , Fatores de Risco
15.
EBioMedicine ; 45: 108-123, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31262715

RESUMO

BACKGROUND: Hydrogen Sulfide (H2S), a third member of gasotransmitter family along with nitric oxide (NO) and carbon monoxide (CO), exerts a wide range of cellular and molecular actions in our body. There is a large body of evidence suggesting that H2S plays an important role in cancer metastasis; however, the molecular mechanisms of H2S-mediated acceleration of cancer metastasis remain unknown. METHODS: We examined the promote effects of H2S on phenotype of gastric cancer (GC) cells (including those of express wild type CD36 and mutant CD36) in vitro and in vivo. GC patients' samples were used for clinical translational significance evaluation. FINDINGS: H2S triggered lipid metabolism reprogramming by significantly up-regulating the expression of the fatty-acid receptor CD36 (CD36) and directly activating CD36 in GC cells. Mechanistically, a disulfide bond located between cysteine (Cys)333 and Cys272 within the CD36 protein structure that was labile to H2S-mediated modification. The long chain-fatty acid (LC-FA) binding pocket was capped by a turn in the CD36 protein, located between helical and sheet structures that were stabilized by the Cys333-Cys272. This limited the secondary binding between LC-FAs and lysine (Lys)334. Breaking the Cys333-Cys272 disulfide bond restored the second LC-FA binding conformation of CD36. Targeting CD36 in vivo blocked H2S-promoted metastasis and improved animal survival. INTERPRETATION: These findings identify that the Cys333-Cys272 disulfide bond disrupted the integrity of the second LC-FA binding conformation of CD36. Therefore, CD36 can directly activate LC-FA access to the cytoplasm by acting as a direct target molecule for H2S.


Assuntos
Antígenos CD36/genética , Proliferação de Células/genética , Cisteína/genética , Neoplasias Gástricas/metabolismo , Animais , Antígenos CD36/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Cisteína/metabolismo , Dissulfetos/metabolismo , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Xenoenxertos , Humanos , Sulfeto de Hidrogênio/metabolismo , Camundongos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Metástase Neoplásica , Óxido Nítrico/genética , Domínios Proteicos/genética , Receptores Odorantes/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia
16.
Life Sci ; 231: 116509, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31152812

RESUMO

Non-coding RNAs (NcRNAs), a family of functional RNA molecules that cannot translate into proteins but control specific gene expression programs, have been shown to be implicated in various biological processes, including fatty acid metabolism. Fast-growing tumor cells rewire their fatty acid metabolic circuitry in order to meet the needs of energy storage, membrane proliferation, and the generation of signaling molecules, which is achieved by regulating a variety of key enzymes along with related signaling pathways in fatty acid metabolism. This review presents an update of our knowledge about the regulatory network of ncRNAs-specifically, microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs)-in this metabolic shift and discusses the possibility of ncRNA-based therapeutics being applied to the restoration of cancer-related fatty acid metabolism.


Assuntos
Ácidos Graxos/metabolismo , RNA não Traduzido/genética , RNA não Traduzido/metabolismo , Reprogramação Celular/fisiologia , Ácidos Graxos/genética , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias/genética , Neoplasias/metabolismo , RNA/genética , RNA/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Transdução de Sinais
17.
Meat Sci ; 156: 75-84, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31132591

RESUMO

The objective of the study was to test the associations between backfat fatty acid composition (FAC) in a sample of 536 Italian Large White pigs and single nucleotide polymorphisms (SNPs) located in candidate genes, using univariate and multivariate approaches. The strongest associations were identified for the SNP AY183428 c.265T>C in Fatty acid synthase (FASN) gene, with the T allele linked to lower backfat contents of stearic (P = 0.003) and arachidic (P < 0.0001) acids, and increased amounts of dihomo-γ-linolenic (P = 0.003) and arachidonic (P = 0.009) acids. These associations were in agreement with the results of a multivariate analysis performed on backfat FAC, and an in silico analysis of the sequence flanking FASN SNP suggested that the T allele may disrupt a putative exonic splicing enhancer sequence therefore possibly affecting FASN activity. If the results will be further confirmed, the studied FASN SNP could be of particular interest for better understanding the regulative patterns controlling porcine backfat FAC.


Assuntos
Ácido Graxo Sintases/genética , Ácidos Graxos/análise , Gordura Subcutânea/química , Sus scrofa/genética , Animais , Ácidos Graxos/genética , Feminino , Masculino , Polimorfismo de Nucleotídeo Único
18.
Methods Mol Biol ; 1978: 259-268, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31119668

RESUMO

Blood vessels are lined by a streamlined monolayer of quiescent endothelial cells (ECs). Although these cells can remain quiescent for years, different stimuli (ischemia, inflammation) and growth factors can activate them and drive a process of new vessel formation (angiogenesis). Emerging evidence reveals that cellular metabolism is a key determinant of the EC subtype specification. The use of stable isotope tracing and mass spectrometry analysis has been essential for the discovery that fatty acid metabolism contributes to EC proliferation and lymphatic EC differentiation. This chapter describes the methodology for setting up palmitate-based tracer metabolomics and the subsequent liquid chromatography-mass spectrometry (LC-MS)-based analysis. As such, tracer metabolomics can be used: (1) to identify the different metabolic pathways relying on carbons provided by fatty acid oxidation and (2) to quantify the relative contributions of palmitate-derived carbons. We begin by providing a background and general principles regarding the use of stable isotopes to study fatty acid metabolism. We then proceed with detailed procedures for the labeling conditions, sample preparation, and subsequent LC-MS analysis.


Assuntos
Cromatografia Líquida/métodos , Ácidos Graxos/metabolismo , Metabolômica/métodos , Espectrometria de Massas em Tandem/métodos , Diferenciação Celular/genética , Células Endoteliais/metabolismo , Ácidos Graxos/genética , Metabolismo dos Lipídeos/genética , Redes e Vias Metabólicas/genética , Palmitatos/química
19.
PLoS One ; 14(5): e0216110, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31120904

RESUMO

BACKGROUND: Genome-wide association studies of common diseases or metabolite quantitative traits often identify common variants of small effect size, which may contribute to phenotypes by modulation of gene expression. Thus, there is growing demand for cellular models enabling to assess the impact of gene regulatory variants with moderate effects on gene expression. Mitochondrial fatty acid oxidation is an important energy metabolism pathway. Common noncoding acyl-CoA dehydrogenase short chain (ACADS) gene variants are associated with plasma C4-acylcarnitine levels and allele-specific modulation of ACADS expression may contribute to the observed phenotype. METHODS AND FINDINGS: We assessed ACADS expression and intracellular acylcarnitine levels in human lymphoblastoid cell lines (LCL) genotyped for a common ACADS variant associated with plasma C4-acylcarnitine and found a significant genotype-dependent decrease of ACADS mRNA and protein. Next, we modelled gradual decrease of ACADS expression using a tetracycline-regulated shRNA-knockdown of ACADS in Huh7 hepatocytes, a cell line with high fatty acid oxidation-(FAO)-capacity. Assessing acylcarnitine flux in both models, we found increased C4-acylcarnitine levels with decreased ACADS expression levels. Moreover, assessing time-dependent changes of acylcarnitine levels in shRNA-hepatocytes with altered ACADS expression levels revealed an unexpected effect on long- and medium-chain fatty acid intermediates. CONCLUSIONS: Both, genotyped LCL and regulated shRNA-knockdown are valuable tools to model moderate, gradual gene-regulatory effects of common variants on cellular phenotypes. Decreasing ACADS expression levels modulate short and surprisingly also long/medium chain acylcarnitines, and may contribute to increased plasma acylcarnitine levels.


Assuntos
Acil-CoA Desidrogenase/genética , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Variação Genética/genética , Acil-CoA Desidrogenase/metabolismo , Carnitina/análogos & derivados , Carnitina/genética , Carnitina/metabolismo , Linhagem Celular Tumoral , Feminino , Estudo de Associação Genômica Ampla/métodos , Genótipo , Hepatócitos/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Mitocôndrias/genética , Mitocôndrias/metabolismo , Oxirredução , Fenótipo
20.
Mol Cell ; 74(3): 598-608.e6, 2019 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-31051140

RESUMO

RNA flow between organisms has been documented within and among different kingdoms of life. Recently, we demonstrated horizontal RNA transfer between honeybees involving secretion and ingestion of worker and royal jellies. However, how the jelly facilitates transfer of RNA is still unknown. Here, we show that worker and royal jellies harbor robust RNA-binding activity. We report that a highly abundant jelly component, major royal jelly protein 3 (MRJP-3), acts as an extracellular non-sequence-specific RNA-aggregating factor. Multivalent RNA binding stimulates higher-order assembly of MRJP-3 into extracellular ribonucleoprotein granules that protect RNA from degradation and enhance RNA bioavailability. These findings reveal that honeybees have evolved a secreted dietary RNA-binding factor to concentrate, stabilize, and share RNA among individuals. Our work identifies high-order ribonucleoprotein assemblies with functions outside cells and organisms.


Assuntos
Abelhas/genética , Ácidos Graxos/genética , Transferência Genética Horizontal/genética , Glicoproteínas/genética , Proteínas de Insetos/genética , Animais , Ácidos Graxos/biossíntese , Transição de Fase , RNA/genética , Transporte de RNA/genética , Proteínas de Ligação a RNA/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA