Project description:The purpose of this study was to identify the changes in gene expression that occur in TWNT-4 human hepatic stellate cells in response to depletion of mannose phosphate isomerase enzymatic activity.
Project description:The purpose of this study was to identify the changes in gene expression that occur in LX-2 human hepatic stellate cells in response to depletion of mannose phosphate isomerase enzymatic activity.
Project description:Bacillus subtilis phosphorylates sugars during or after their transport into the cell.Perturbation in the conversion of intracellular phosphosugars to the central carbon metabolitesand accumulation of phosphosugars can impose stress on the cells. In this study, we investigated the effect of phosphosugar stress on B. subtilis. Preliminary experiments indicated that the non-matabolizable analogs of glucose were unable to impose stress on B. subtilis. In contrast, deletion of manA encoding mannose 6-phosphate isomerase (responsible for conversion of mannose 6-phosphate to fructose 6-phosphate) resulted in growth arrest and bulged cell shape in the medium containing mannose. Besides, an operon encoding a repressor (GlcR) and a haloic acid dehalogenase (HAD)-like phosphatase (PhoC; previously YwpJ) were upregulated. Integration of the PglcR-lacZ cassette into different mutational backgrounds indicated that PglcR is induced when (i) a manA-deficient strain is cultured with mannose or (ii) when glcR is deleted.GlcR represses the transcription of glcR-phoC bybinding to the A-type core elements of PglcR. Electrophoretic mobility shift assay showed no interaction between mannose 6-phosphate (or other phosphosugars) and the GlcR-PglcR DNA complex. PhoCwas an acid phosphatase mainly able to dephosphorylate glycerol 3-phosphate and ribose 5- phosphate. Mannose 6-phosphatewas only weakly dephosphorylated by PhoC. Since deletion of glcR and phoCalone or in combination had no effect on the cell duringphosphosugar stress,it is assumed that the derepression of glcR-phoC is a side effect of phosphosugar stress in B. subtilis.
Project description:Bacillus subtilis has different response systems to cope with external and internal stressors. In this study, we investigated the effect of phosphosugar stress caused by accumulation of phosphosugars in B. subtilis. To do so, manA, the encoding gene of mannose-6-phosphate isomerase, was deleted in B. subtilis KM0 to construct strain KM642. Next, strains KM0 and KM642 were cultured in LB with 1% mannose and the cell pellets were isolated after 3.5 h of incubation at 37 °C for transcriptome analysis by RNA-Seq. Differential gene expression analysis was performed based on the RNA-Seq data generated by paired end sequencing (2 x 75nt read length) of Illumina TruSeq stranded mRNA-cDNA libraries on Illumina MiSeq system from control strain and manA deletion mutant.
Project description:This study explored the regulatory effect of D-mannose on host immunometabolic responses after viral infection. The results showed that D-mannose can compete with glucose for transporters and hexokinase, inhibiting glycolysis, reducing mitochondrial reactive oxygen species and succinate-induced HIF-1α, thereby reducing virus-induced inflammatory cytokine production. Even with delayed combinatorial treatment of D-mannose and antiviral monotherapy after viral infection, a synergistic effect was still observed in mouse models. Phosphomannose isomerase (PMI) activity determines the benefits of D-mannose, as simultaneous PMI depletion and mannose supplementation impaired cell viability. PMI inhibition can suppress replication of various viruses by affecting host and viral surface protein glycosylation. However, D-mannose does not inhibit PMI activity or viral fitness. In summary, PMI-centered therapeutic strategies can eliminate viral infections, while D-mannose treatment reprograms glycolysis to control collateral damage.
Project description:The target proteins phosphoglycerate kinase 2 (PGK2), glycerol-3-phosphate dehydrogenase (GPD2)GPD2 and glucose-6-phosphate isomerase (GPI) were screened by combining transcriptome, proteomics and reverse docking We detected the binding constant of the active compound using microscale thermophoresis (MST). It was found that esculetin bound well with three potential target proteins.
Project description:We used RNA-seq to profile E. coli K-12 MG1655 strains subjected to adaptive laboratory evolution after knockout of endogenous glucose-6-phosphate isomerase (pgi) and subsequent expression of heterologous version of the pgi gene from Pseudomonas aeruginosa and Bacillus megaterium.
Project description:- Identification of proteins whose expression was affected by a putative glucose 6-phosphate isomerase in Acidovorax citrulli str. KACC17005 - Shotgun proteomic analysis was used - Two strains were used with three biological replicates (total 6 samples). WT_4C: the wild-type strain. 4C: 2, a putative glucose 6-phosphate isomerase knockout mutant
Project description:We have repurposed standard phosphoproteomics workflow based on Fe3+IMAC for enrichment ofmannose-6-phosphate modified glycopeptides. This worklfow was used to profile lysosomal acid hydrolases in HeLa and CHO cell lines. We have combined this approach with CRISPR/Cas9 KO of acid phosphatases 2 and 5 responsible for dephosphorylation of mannose-6-phosphate glycopeptides in the lysosome. This combined approach enabled significantly deeper coverage of CHO mannose-6-phosphate glycoproteome.
