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Mutations in the Drosophila melanogaster gene pyruvate kinase interfere with larval development

ABSTRACT: Mutations in the Drosophila melanogaster gene pyruvate kinase interfere with larval development

PROVIDER: PRJNA980708 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24843639_1.fastq.gz	Fastqsanger.gz
	SRR24843639_2.fastq.gz	Fastqsanger.gz
	SRR24843640_1.fastq.gz	Fastqsanger.gz
	SRR24843640_2.fastq.gz	Fastqsanger.gz
	SRR24843641_1.fastq.gz	Fastqsanger.gz

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Mutations in the Drosophila melanogaster gene pyruvate kinase interfere with larval development

Project description:Pyruvate kinase (Pyk) is a rate-limiting enzyme that catalyzes the final metabolic reaction in glycolysis. The importance of this enzyme, however, extends far beyond ATP production, as Pyk is also known to regulate tissue growth, cell proliferation, and development. Studies of this enzyme in Drosophila melanogaster, however, are complicated by the fact the fly genome encodes six Pyk paralogs and the function of these individual genes remain poorly defined. To address this issue, we used a phylogenetic approach to demonstrate that the gene Pyk encodes the enzyme most similar to the mammalian Pyk orthologs while the other five Drosophila Pyk have significantly diverged from the canonical enzyme. Consistent with this observation, metabolomic studies of two different Pyk mutant backgrounds revealed that larvae lacking Pyk exhibit a severe block in glycolysis, with a buildup of glycolytic intermediates upstream of pyruvate. However, our analysis also unexpectantly reveals that steady state pyruvate levels are unchanged in Pyk mutants, indicating that larval metabolism maintains pyruvate pool size despite severe metabolic limitations. Consistent with our metabolomics findings, a complementary RNAseq analysis revealed that genes involved in lipid metabolism and peptidase activity are elevated in Pyk mutants, again indicating that loss of this glycolytic enzyme induces compensatory changes in other aspects of metabolism. Overall, our study provides both insight towards understanding how Drosophila larval metabolism adapts to disruption of glycolytic metabolism and immediate clinical relevance considering that Pyk deficiency is the most common congenital enzymatic defect in humans.

2023-09-01 | GSE234299 | GEO

Gene expression profiling of NUAK kinase overexpression in Drosophila larval muscle development

Project description:Signal transduction pathways mediated by kinases control diverse biological outputs at the level of cells and tissues to regulate a diverse array of biological and developmental events. To gain insight into how muscle expression of the evolutionarily conserved NUAK kinase regulates the transcriptional landscape during Drosophila melanogaster development, we performed high-throughput sequencing of RNA from either whole larvae or dissected muscle fillets at the end of larval development. Raw data was generated using the Illumina HiSeq platform. After trimming and mapping to the Drosophila reference genome, differential gene expression and GO enrichment analysis were completed.

2022-05-30 | GSE204894 | GEO

Pharmacological activation of pyruvate kinase M2 (PKM2) inhibits CD4+ T cell pathogenicity and suppresses autoimmunity

Project description:Pyruvate kinase (PK) catalyzes the conversion of phosphoenolpyruvate to pyruvate during glycolysis. The PK isoform PKM2 has additional roles in regulation of gene transcription and protein phosphorylation. PKM2 controls macrophage metabolic remodeling in inflammation, but its role in T cell biology is poorly understood. These results show that TEPP-46, an activator of PKM2, reduces CD4+ T cell activation, proliferation, and cytokine production by inhibiting essential signaling pathways and preventing glycolysis.

2019-11-26 | GSE139378 | GEO

Pyruvate dehydrogenase kinase is a metabolic checkpoint for polarization of macrophages to the M1 phenotype

Project description:Metabolic reprogramming during macrophage polarization supports the effector functions of these cells in health and disease. Although the importance of glycolytic and oxidative metabolism in M1 and M2 macrophages, respectively, is well established, our knowledge of metabolic checkpoints controlling these effector states is limited. Here we demonstrate that pyruvate dehydrogenase kinase (PDK), which inhibits the conversion of cytosolic pyruvate to mitochondrial acetyl-CoA by pyruvate dehydrogenase, functions as a metabolic checkpoint in M1 macrophages. Genetic deletion or pharmacological inhibition of PDK2/4 prevents polarization of macrophages to the M1 phenotype in response to inflammatory stimuli (lipopolysaccharide plus IFN-γ). The therapeutic potential of attenuation of pro-inflammatory responses by PDK inhibition was tested, both genetically and pharmacologically, in obesity-induced insulin resistance, a disease process in which M1 macrophages contribute to adipose tissue inflammation and insulin resistance. Taken together, these studies identify PDK2/4 as a metabolic checkpoint for M1 phenotype polarization of macrophages.

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Haiman2020 - Pyruvate Kinase Regulatory Model of Erythrocyte Glycolysis

Project description:Dynamic model utilizing mass action kinetics to elucidate network-level effects of allosteric regulation in erythrocyte metabolism. The pyruvate kinase regulatory model includes the glycolytic pathway, the Rapaport Leubering (R.L.) Shunt, hemoglobin-oxygen binding, and elementary reaction steps to represent the enzymatic mechanisms for Pyruvate Kinase (PYK). To demonstrate the sampling and ensemble handling capabilities of MASSpy, an eensemble modeling approach is utilized to assess the dynamics of pyruvate kinase regulation. For the ensemble modeling feature demonstration, a range of parameter sets are generated through Markov-chain Monte Carlo (MCMC) sampling to represent the biological variability across the ensemble of models. Transient dynamic behavior and model steady states are studied through dynamic simulation and analysis. All models associated with the ensemble-modeling example are found in the GitHub repository at (https://github.com/SBRG/MASSpy-publication), including iPython notebooks to construct all model variations presented in the publication.

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The NTP Generating Activity of Pyruvate Kinase II is Critical for Apicoplast Maintenance in Plasmodium falciparum

Project description:The apicoplast of Plasmodium falciparum parasites is believed to rely on the import of three-carbon phosphate compounds for use in organelle anabolic pathways, in addition to the generation of energy and reducing power within the organelle. We generated a series of genetic deletions in an apicoplast metabolic bypass line to determine which genes involved in apicoplast carbon metabolism are required for blood-stage parasite survival and organelle maintenance. We were able to demonstrate that pyruvate kinase II (PyrKII), the sole predicted source of pyruvate and NTPs within the apicoplast, is essential for organelle maintenance. However, deletions within apicoplast pathways that metabolize pyruvate did not result in organelle disruption, indicating that NTPs generated by PyrKII are the products required for this process. Enzymatic characterization of PyrKII revealed that it was active against all NDPs and dNDPs tested with broad substrate specificity, suggesting that it may be capable of generating any and all NTPs and dNTPs within the organelle. Conditional mislocalization of PyrKII resulted in a decrease in transcript levels within the apicoplast that preceded organelle disruption, suggesting that PyrKII is required for organelle maintenance due to its role in nucleotide triphosphate generation within the apicoplast.

2020-07-31 | GSE136688 | GEO

Adult and larval tracheal systems exhibit different molecular architectures in Drosophila

Project description:We analysed the RNA-seq profile of larval and adult trachea of the fruitfly Drosophila melanogaster, as well as from whole flies.

2024-02-12 | GSE225000 | GEO

Gene expression profiling of NUAK kinase overexpression in Drosophila larval muscle development

Project description:Gene expression profiling of NUAK kinase overexpression in Drosophila larval muscle development

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Drosophila melanogaster female late third instar larval gonads single-cell RNA-Seq (scRNA-Seq) profiling

Project description:The larval ovary is made up of multiple cell types including germ cells and somatic cells. The diversity of cell types and transcriptional regulation is not fully understood. To get single cell resolution of larval ovary regulation, we generated single-cell RNA expression profiles (scRNA-Seq) from late third instar larval ovaries of a reference Drosophila melanogaster genotype w[1118].

2019-02-01 | GSE125948 | GEO

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2019-03-22 | GSE128646 | GEO

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