In this study we use comparative genomics to uncover a gene with uncharacterized function (1700011H14Rik/C14orf105/CCDC198), which we hereby name FAME (Factor Associated with Metabolism and Energy). We observe that FAME shows an unusually high evolutionary divergence in birds and mammals. Through the comparison of single nucleotide polymorphisms, we identify gene flow of FAME from Neandertals into modern humans. We conduct knockout experiments on animals and observe altered body weight and decreased energy expenditure in Fame knockout animals, corresponding to genome-wide association studies linking FAME with higher body mass index in humans. Gene expression and subcellular localization analyses reveal that FAME is a membrane-bound protein enriched in the kidneys. Although the gene knockout results in structurally normal kidneys, we detect higher albumin in urine and lowered ferritin in the blood. Through experimental validation, we confirm interactions between FAME and ferritin and show co-localization in vesicular and plasma membranes.

• MeSH
• celogenomová asociační studie * MeSH
• energetický metabolismus * genetika   MeSH
• ferritin genetika   MeSH
• ledviny MeSH
• lidé MeSH
• neandertálci MeSH
• tělesná hmotnost MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ferritin MeSH

G-protein-coupled receptor GPR10 is expressed in brain areas regulating energy metabolism. In this study, the effects of GPR10 gene deficiency on energy homeostasis in mice of both sexes fed either standard chow or a high-fat diet (HFD) were studied, with a focus on neuronal activation of PrRP neurons, and adipose tissue and liver metabolism. GPR10 deficiency in males upregulated the phasic and tonic activity of PrRP neurons in the nucleus of the solitary tract. GPR10 knockout (KO) males on a standard diet displayed a higher body weight than their wild-type (WT) littermates due to an increase in adipose tissue mass; however, HFD feeding did not cause weight differences between genotypes. Expression of lipogenesis genes was suppressed in the subcutaneous adipose tissue of GPR10 KO males. In contrast, GPR10 KO females did not differ in body weight from their WT controls, but showed elevated expression of lipid metabolism genes in the liver and subcutaneous adipose tissue compared to WT controls. An attenuated non-esterified fatty acids change after glucose load compared to WT controls suggested a defect in insulin-mediated suppression of lipolysis in GPR10 KO females. Indirect calorimetry did not reveal any differences in energy expenditure among groups. In conclusion, deletion of GPR10 gene resulted in changes in lipid metabolism in mice of both sexes, however in different extent. An increase in adipose tissue mass observed in only GPR10 KO males may have been prevented in GPR10 KO females owing to a compensatory increase in the expression of metabolic genes.

• Klíčová slova
• Energy expenditure, GPR10 KO mice, Gene expression, Neuronal activity, Prolactin-releasing peptide, Standard and high-fat diets,
• MeSH
• energetický metabolismus genetika   MeSH
• homeostáza genetika   MeSH
• hormon uvolňující prolaktin metabolismus   MeSH
• inzulinová rezistence genetika   MeSH
• metabolismus lipidů genetika   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• obezita genetika   MeSH
• receptory spřažené s G-proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hormon uvolňující prolaktin MeSH
• Prlh protein, mouse MeSH Prohlížeč
• Prlhr protein, mouse MeSH Prohlížeč
• receptory spřažené s G-proteiny MeSH

The mitochondrial inner membrane glycerophospholipid cardiolipin (CL) associates with mitochondrial proteins to regulate their activities and facilitate protein complex and supercomplex formation. Loss of CL leads to destabilized respiratory complexes and mitochondrial dysfunction. The role of CL in an organism lacking a conventional electron transport chain (ETC) has not been elucidated. Trypanosoma brucei bloodstream forms use an unconventional ETC composed of glycerol-3-phosphate dehydrogenase and alternative oxidase (AOX), while the mitochondrial membrane potential (ΔΨm) is generated by the hydrolytic action of the Fo F1 -ATP synthase (aka Fo F1 -ATPase). We now report that the inducible depletion of cardiolipin synthase (TbCls) is essential for survival of T brucei bloodstream forms. Loss of CL caused a rapid drop in ATP levels and a decline in the ΔΨm. Unbiased proteomic analyses revealed a reduction in the levels of many mitochondrial proteins, most notably of Fo F1 -ATPase subunits and AOX, resulting in a strong decline of glycerol-3-phosphate-stimulated oxygen consumption. The changes in cellular respiration preceded the observed decrease in Fo F1 -ATPase stability, suggesting that the AOX-mediated ETC is the first pathway responding to the decline in CL. Select proteins and pathways involved in glucose and amino acid metabolism were upregulated to counteract the CL depletion-induced drop in cellular ATP.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• elektronový transportní řetězec metabolismus   MeSH
• energetický metabolismus genetika   MeSH
• geneticky modifikované organismy MeSH
• genový knockout * MeSH
• glycerolfosfátdehydrogenasa metabolismus   MeSH
• kardiolipiny genetika   metabolismus   MeSH
• membránové proteiny genetika   metabolismus   MeSH
• membránový potenciál mitochondrií genetika   MeSH
• mitochondriální membrány metabolismus   MeSH
• mitochondriální proteiny metabolismus   MeSH
• mitochondrie metabolismus   MeSH
• oxidoreduktasy metabolismus   MeSH
• proteom MeSH
• proteomika MeSH
• protozoální proteiny genetika   metabolismus   MeSH
• rostlinné proteiny metabolismus   MeSH
• spotřeba kyslíku genetika   MeSH
• transferasy pro jiné substituované fosfátové skupiny genetika   metabolismus   MeSH
• Trypanosoma brucei brucei klasifikace   genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenosintrifosfát MeSH
• alternative oxidase MeSH Prohlížeč
• cardiolipin synthetase MeSH Prohlížeč
• elektronový transportní řetězec MeSH
• glycerolfosfátdehydrogenasa MeSH
• kardiolipiny MeSH
• membránové proteiny MeSH
• mitochondriální proteiny MeSH
• oxidoreduktasy MeSH
• proteom MeSH
• protozoální proteiny MeSH
• rostlinné proteiny MeSH
• transferasy pro jiné substituované fosfátové skupiny MeSH

We report results showing that the silencing of carbonic anhydrase I (siCA1) in prostatic (PC3) tumour cells has a significant impact on exosome formation. An increased diameter, concentration and diversity of the produced exosomes were noticed as a consequence of this knock-down. The protein composition of the exosomes' cargo was also altered. Liquid chromatography and mass spectrometry analyses identified 42 proteins significantly altered in PC3 siCA1 exosomes compared with controls. The affected proteins are mainly involved in metabolic processes, biogenesis, cell component organization and defense/immunity. Interestingly, almost all of them have been described as 'enhancers' of tumour development through the promotion of cell proliferation, migration and invasion. Thus, our results indicate that the reduced expression of the CA1 protein enhances the malignant potential of PC3 cells.

• Klíčová slova
• LC-MS, PC3 cells, carbonic anhydrase I, exosomes, malignant potential, siCA1, siMock,
• MeSH
• buňky PC-3 MeSH
• energetický metabolismus genetika   MeSH
• exozómy genetika   metabolismus   MeSH
• karboanhydrasa I genetika   metabolismus   MeSH
• lidé MeSH
• nádory prostaty genetika   metabolismus   patologie   MeSH
• pohyb buněk genetika   MeSH
• proliferace buněk genetika   MeSH
• regulace genové exprese enzymů * MeSH
• regulace genové exprese u nádorů * MeSH
• RNA interference * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• karboanhydrasa I MeSH

Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide. Among the PPV strains identified so far, only PPV-C, PPV-CR, and PPV-CV are able to infect cherries under natural conditions. Herein, we evaluated the pathogenic potential of two viral isolates in herbaceous host Nicotiana benthamiana. Significantly higher accumulation of PPV capsid protein in tobacco leaves infected with PPV-CR (RU-30sc isolate) was detected in contrast to PPV-C (BY-101 isolate). This result correlated well with the symptoms observed in the infected plants. To further explore the host response upon viral infection at the molecular level, a comprehensive proteomic profiling was performed. Using reverse-phase ultra-high-performance liquid chromatography followed by label-free mass spectrometry quantification, we identified 38 unique plant proteins as significantly altered due to the infection. Notably, the abundances of photosynthesis-related proteins, mainly from the Calvin-Benson cycle, were found more aggressively affected in plants infected with PPV-CR isolate than those of PPV-C. This observation was accompanied by a significant reduction in the amount of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of proteins that are involved in stimulation of photosynthetic capacity, modification of amino acid, and carbohydrate metabolism may affect plant growth and initiate energy formation via gluconeogenesis in PPV infected N. benthamiana. Furthermore, we suggest that the higher accumulation of H2O2 in PPV-CR infected leaves plays a crucial role in plant defense and development by activating the glutathione synthesis.

• Klíčová slova
• LC−MS, antioxidants, compatible interaction, label-free quantification, photosynthesis, sharka, tobacco,
• MeSH
• chlorofyl biosyntéza   MeSH
• chromatografie s reverzní fází MeSH
• energetický metabolismus genetika   MeSH
• fotosyntéza genetika   MeSH
• genotyp MeSH
• glutathion biosyntéza   MeSH
• hmotnostní spektrometrie MeSH
• interakce hostitele a patogenu genetika   MeSH
• karotenoidy biosyntéza   MeSH
• listy rostlin genetika   metabolismus   virologie   MeSH
• nemoci rostlin genetika   virologie   MeSH
• oxidace-redukce MeSH
• peroxid vodíku metabolismus   MeSH
• proteiny tepelného šoku klasifikace   genetika   metabolismus   MeSH
• Prunus avium virologie   MeSH
• regulace genové exprese u rostlin * MeSH
• rostlinné proteiny klasifikace   genetika   metabolismus   MeSH
• slivoň švestka virologie   MeSH
• tabák genetika   metabolismus   virologie   MeSH
• virus šarky švestky klasifikace   genetika   růst a vývoj   patogenita   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chlorofyl MeSH
• glutathion MeSH
• karotenoidy MeSH
• peroxid vodíku MeSH
• proteiny tepelného šoku MeSH
• rostlinné proteiny MeSH

Brown adipose tissue (BAT) plays an important role in lipid and glucose metabolism in rodents and possibly also in humans. Identification of genes responsible for BAT function would shed light on underlying pathophysiological mechanisms of metabolic disturbances. Recent linkage analysis in the BXH/HXB recombinant inbred (RI) strains, derived from Brown Norway (BN) and spontaneously hypertensive rats (SHR), identified two closely linked quantitative trait loci (QTL) associated with glucose oxidation and glucose incorporation into BAT lipids in the vicinity of Wars2 (tryptophanyl tRNA synthetase 2 (mitochondrial)) gene on chromosome 2. The SHR harbors L53F WARS2 protein variant that was associated with reduced angiogenesis and Wars2 thus represents a prominent positional candidate gene. In the current study, we validated this candidate as a quantitative trait gene (QTG) using transgenic rescue experiment. SHR-Wars2 transgenic rats with wild type Wars2 gene when compared to SHR, showed more efficient mitochondrial proteosynthesis and increased mitochondrial respiration, which was associated with increased glucose oxidation and incorporation into BAT lipids, and with reduced weight of visceral fat. Correlation analyses in RI strains showed that increased activity of BAT was associated with amelioration of insulin resistance in muscle and white adipose tissue. In summary, these results demonstrate important role of Wars2 gene in regulating BAT function and consequently lipid and glucose metabolism.

• MeSH
• energetický metabolismus * genetika   MeSH
• fenotyp MeSH
• genetická predispozice k nemoci MeSH
• genetické asociační studie MeSH
• glukosa metabolismus   MeSH
• hnědá tuková tkáň metabolismus   patologie   MeSH
• kultivované buňky MeSH
• lokus kvantitativního znaku MeSH
• metabolismus lipidů MeSH
• mitochondrie metabolismus   MeSH
• mutace * MeSH
• nitrobřišní tuk metabolismus   patofyziologie   MeSH
• obezita genetika   metabolismus   patofyziologie   MeSH
• potkani inbrední SHR MeSH
• tryptofan-tRNA-ligasa genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glukosa MeSH
• tryptofan-tRNA-ligasa MeSH

We previously mapped hypertension-related insulin resistance quantitative trait loci (QTLs) to rat chromosomes 4, 12 and 16 using adipocytes from F2 crosses between spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats, and subsequently identified Cd36 as the gene underlying the chromosome 4 locus. The identity of the chromosome 12 and 16 genes remains unknown. To identify whole-body phenotypes associated with the chromosome 12 and 16 linkage regions, we generated and characterised new congenic strains, with WKY donor segments introgressed onto an SHR genetic background, for the chromosome 12 and 16 linkage regions. We found a >50% increase in insulin sensitivity in both the chromosome 12 and 16 strains. Blood pressure and left ventricular mass were reduced in the two congenic strains consistent with the congenic segments harbouring SHR genes for insulin resistance, hypertension and cardiac hypertrophy. Integrated genomic analysis, using physiological and whole-genome sequence data across 42 rat strains, identified variants within the congenic regions in Upk3bl, RGD1565131 and AABR06087018.1 that were associated with blood pressure, cardiac mass and insulin sensitivity. Quantitative trait transcript analysis across 29 recombinant inbred strains showed correlation between expression of Hspb1, Zkscan5 and Pdgfrl with adipocyte volume, systolic blood pressure and cardiac mass, respectively. Comparative genome analysis showed a marked enrichment of orthologues for human GWAS-associated genes for insulin resistance within the syntenic regions of both the chromosome 12 and 16 congenic intervals. Our study defines whole-body phenotypes associated with the SHR chromosome 12 and 16 insulin-resistance QTLs, identifies candidate genes for these SHR QTLs and finds human orthologues of rat genes in these regions that associate with related human traits. Further study of these genes in the congenic strains will lead to robust identification of the underlying genes and cellular mechanisms.

• Klíčová slova
• Congenic, Genomic, Hypertension, Insulin resistance, Rat,
• MeSH
• celogenomová asociační studie MeSH
• energetický metabolismus genetika   MeSH
• genomika * MeSH
• homeostáza MeSH
• hypertenze genetika   patofyziologie   MeSH
• inzulin farmakologie   MeSH
• inzulinová rezistence genetika   MeSH
• játra účinky léků   metabolismus   MeSH
• jednonukleotidový polymorfismus genetika   MeSH
• kalorimetrie MeSH
• kardiomegalie genetika   patofyziologie   MeSH
• kosterní svaly účinky léků   metabolismus   MeSH
• krevní tlak účinky léků   MeSH
• lidé MeSH
• lokus kvantitativního znaku genetika   MeSH
• potkani inbrední SHR MeSH
• regulace genové exprese účinky léků   MeSH
• savčí chromozomy genetika   MeSH
• srdeční komory účinky léků   patologie   MeSH
• stravovací zvyklosti účinky léků   MeSH
• tělesná hmotnost účinky léků   MeSH
• triglyceridy metabolismus   MeSH
• velikost orgánu účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• inzulin MeSH
• triglyceridy MeSH

Many high-quality genomes are available for dixenous (two hosts) trypanosomatid species of the genera Trypanosoma, Leishmania, and Phytomonas, but only fragmentary information is available for monoxenous (single-host) trypanosomatids. In trypanosomatids, monoxeny is ancestral to dixeny, thus it is anticipated that the genome sequences of the key monoxenous parasites will be instrumental for both understanding the origin of parasitism and the evolution of dixeny. Here, we present a high-quality genome for Leptomonas pyrrhocoris, which is closely related to the dixenous genus Leishmania. The L. pyrrhocoris genome (30.4 Mbp in 60 scaffolds) encodes 10,148 genes. Using the L. pyrrhocoris genome, we pinpointed genes gained in Leishmania. Among those genes, 20 genes with unknown function had expression patterns in the Leishmania mexicana life cycle suggesting their involvement in virulence. By combining differential expression data for L. mexicana, L. major and Leptomonas seymouri, we have identified several additional proteins potentially involved in virulence, including SpoU methylase and U3 small nucleolar ribonucleoprotein IMP3. The population genetics of L. pyrrhocoris was also addressed by sequencing thirteen strains of different geographic origin, allowing the identification of 1,318 genes under positive selection. This set of genes was significantly enriched in components of the cytoskeleton and the flagellum.

• MeSH
• druhová specificita MeSH
• energetický metabolismus genetika   MeSH
• fylogeneze MeSH
• genom protozoální genetika   MeSH
• genová ontologie MeSH
• Leishmania klasifikace   genetika   patogenita   MeSH
• molekulární evoluce * MeSH
• protozoální geny genetika   MeSH
• stanovení celkové genové exprese metody   MeSH
• Trypanosomatina klasifikace   genetika   patogenita   MeSH
• virulence genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Glycolytic shift is a characteristic feature of rapidly proliferating cells, such as cells during development and during immune response or cancer cells, as well as of stem cells. It results in increased glycolysis uncoupled from mitochondrial respiration, also known as the Warburg effect. Notch signalling is active in contexts where cells undergo glycolytic shift. We decided to test whether metabolic genes are direct transcriptional targets of Notch signalling and whether upregulation of metabolic genes can help Notch to induce tissue growth under physiological conditions and in conditions of Notch-induced hyperplasia. We show that genes mediating cellular metabolic changes towards the Warburg effect are direct transcriptional targets of Notch signalling. They include genes encoding proteins involved in glucose uptake, glycolysis, lactate to pyruvate conversion and repression of the tricarboxylic acid cycle. The direct transcriptional upregulation of metabolic genes is PI3K/Akt independent and occurs not only in cells with overactivated Notch but also in cells with endogenous levels of Notch signalling and in vivo. Even a short pulse of Notch activity is able to elicit long-lasting metabolic changes resembling the Warburg effect. Loss of Notch signalling in Drosophila wing discs as well as in human microvascular cells leads to downregulation of glycolytic genes. Notch-driven tissue overgrowth can be rescued by downregulation of genes for glucose metabolism. Notch activity is able to support growth of wing during nutrient-deprivation conditions, independent of the growth of the rest of the body. Notch is active in situations that involve metabolic reprogramming, and the direct regulation of metabolic genes may be a common mechanism that helps Notch to exert its effects in target tissues.

• Klíčová slova
• Notch targets, Warburg effect, glycolytic shift, metabolism, tissue growth,
• MeSH
• aktivace transkripce MeSH
• biologické modely MeSH
• buněčné linie MeSH
• citrátový cyklus genetika   MeSH
• energetický metabolismus genetika   MeSH
• exprese genu MeSH
• glykolýza genetika   MeSH
• lidé MeSH
• promotorové oblasti (genetika) MeSH
• proteiny Drosophily genetika   metabolismus   MeSH
• receptory Notch genetika   metabolismus   MeSH
• regulace genové exprese * MeSH
• regulační oblasti nukleových kyselin MeSH
• reportérové geny MeSH
• represorové proteiny genetika   metabolismus   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• proteiny Drosophily MeSH
• receptory Notch MeSH
• represorové proteiny MeSH
• Su(H) protein, Drosophila MeSH Prohlížeč

Pseudomonas fluorescens is a well-known food spoiler, able to cause serious economic losses in the food industry due to its ability to produce many extracellular, and often thermostable, compounds. The most outstanding spoilage events involving P. fluorescens were blue discoloration of several food stuffs, mainly dairy products. The bacteria involved in such high-profile cases have been identified as belonging to a clearly distinct phylogenetic cluster of the P. fluorescens group. Although the blue pigment has recently been investigated in several studies, the biosynthetic pathway leading to the pigment formation, as well as its chemical nature, remain challenging and unsolved points. In the present paper, genomic and transcriptomic data of 4 P. fluorescens strains (2 blue-pigmenting strains and 2 non-pigmenting strains) were analyzed to evaluate the presence and the expression of blue strain-specific genes. In particular, the pangenome analysis showed the presence in the blue-pigmenting strains of two copies of genes involved in the tryptophan biosynthesis pathway (including trpABCDF). The global expression profiling of blue-pigmenting strains versus non-pigmenting strains showed a general up-regulation of genes involved in iron uptake and a down-regulation of genes involved in primary metabolism. Chromogenic reaction of the blue-pigmenting bacterial cells with Kovac's reagent indicated an indole-derivative as the precursor of the blue pigment. Finally, solubility tests and MALDI-TOF mass spectrometry analysis of the isolated pigment suggested that its molecular structure is very probably a hydrophobic indigo analog.

• Klíčová slova
• Blue mozzarella, Comparative genomics and transcriptomics, Pseudomonas fluorescens,
• MeSH
• biologické pigmenty genetika   MeSH
• down regulace MeSH
• energetický metabolismus genetika   MeSH
• fenotyp MeSH
• fylogeneze MeSH
• genomika MeSH
• mléčné výrobky mikrobiologie   MeSH
• oxidoreduktasy genetika   MeSH
• potravinářská mikrobiologie * MeSH
• Pseudomonas fluorescens genetika   metabolismus   MeSH
• spotřeba kyslíku genetika   MeSH
• stanovení celkové genové exprese MeSH
• transkriptom genetika   MeSH
• tryptofan biosyntéza   MeSH
• upregulace MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické pigmenty MeSH
• oxidoreduktasy MeSH
• tryptofan MeSH