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ABSTRACT:
PROVIDER: PRJNA1223443 | ENA |
REPOSITORIES: ENA
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Project description:UDP-glucuronic acid (UDP-GlcUA) is a nucleotide sugar that plays important roles in many organisms and excessive UDP-GlcUA in the cell causes many defects in the cellular processes. In Cryptococcus spp., mutations in the UXS1 gene which encodes an enzyme that converts UDP-GlcUA into UDP-xylose trigger high level accumulation of UDP-GlcUA and effectuate resistance to the antifungal drug 5-fluorocytosine. Here, we show that elevation of UDP-GlcUA affects several cellular processes including growth rate, ability to grow at various stress conditions, and resistance to fluorine containing analogs. RNA-seq analyses reveal that UXS1 deletion leads to the identification of three differentially expressed endopeptidase genes, notably PEP401. Lack of PEP401 from the uxs1 mutant background reduces the UDP-GlcUA levels and reverts all the phenotypes of the uxs1 mutant toward the wild-type characteristics. Particularly, high levels of UDP-GlcUA not only regulate expression of PEP401 at RNA and protein levels, but it also enhances the proteolytic activity of total protein extracts in a PEP401-dependent manner, establishing a functional link between nucleotide sugar metabolism and proteolytic regulation. Moreover, the UDP-GlcUA transporter gene, UUT1, can further modulate the levels of UDP-GlcUA in the uxs1 pep401 double mutant and manifests the drug resistance phenotypes observed in the uxs1 mutant. Taken together, these findings reveal a previously unrecognized regulatory network that connects UDP-GlcUA metabolism to protease-mediated cellular processes and to the transportation of UDP-GlcUA. This newly identified interplay provides a foundation for targeting nucleotide sugar metabolism and protease regulation in the development of improved therapeutic strategies against cryptococcosis.
2025-04-08 | GSE289668 | GEO
Project description:Nucleotide sugar transporters (NSTs) are ER and Golgi-resident members of the solute carrier 35 (SLC35) family which supply substrates for glycosylation by exchanging lumenal nucleotide monophosphates for cytosolic nucleotide sugars. Defective NSTs have been associated with congenital disorders of glycosylation (CDG), however, molecular basis of many types of CDG remains poorly characterized. To better understand CDG biology, we identified potential interaction partners of UDP-galactose transporter (SLC35A2), UDP-N-acetylglucosamine transporter (SLC35A3) and an orphan nucleotide sugar transporter SLC35A4. For this purpose, each of the SLC35A2-A4 proteins was used as a bait in four independent pull-down experiments and the identity of the immunoprecipitated material was discovered using MS techniques. The consensus findings for each of the NSTs tested were listed as potential interaction partners and should prove useful as a starting point for deciphering the NST interaction network.
2020-12-21 | MSV000086622 | MassIVE
Project description:Glycosylation reactions require activated glycosyl donors in form of nucleotide sugars to drive processes such as post-translational protein modifications, glycolipid and polysaccharide biosynthesis. Most of these reactions occur in the Golgi requiring cytosolic-derived nucleotide sugars, which are actively transferred into the Golgi lumen by nucleotide sugar transporters. Here we present the identification of the plant UDP-N-acetylglucosamine (UDP-GlcNAc) transporter (UGNT1) indispensable for the delivery of a substrate for maturation of N-glycans and glycosyl inositol phosphorylceramides (GIPCs). Profiles of N-glycopeptides revealed that UGNT1 loss-of-function mutants are devoid of complex and hybrid N-glycans. Instead, most of the glycol-N-peptide population contained high mannose structures, representing the structure prior to the addition of the first GlcNAc in the Golgi. Our findings emphasize that the reference plant Arabidopsis contains a single UDP-GlcNAc transporter responsible for the maturation of complex N-glycans in the Golgi lumen.
2018-09-24 | PXD006635 | Pride
Project description:Our previous investigation indicated that high-virulence C. gattii (C. gattii TIMM 4097) tend to reside in the alveoli, whereas low-virulence C. gattii (C. gattii TIMM 4903) tend to be washed out from the alveoli and move into the central side of the respiratory system. To test this hypothesis, we performed microarray assay. C. gattii TIMM 4097 and C. gattii TIMM 4903 were prepared for the study. The mice were anesthetized and the spore suspension was intratracheally injected into each mouse. Mice were sacrificed 15 days after the infection and then examined. Lung of the mice infected with C. gattii TIMM 4097 and C. gattii TIMM 4903 and normal controls were obtained (n=3, respectively. RNA targets preparation was performed according to the manufacturer's protocol using GeneChip(R) 3' IVT Express Kit (Affymetrix). One hundred nanograms of total RNA were converted into double-stranded cDNA template for transcription. In vitro transcription synthesized amplified RNA (aRNA) and incorporated a biotin-conjugated nucleotide. After purification and fragmentation of aRNA, 12.5 ug of them was hybridized to GeneChip(R) Mouse Genome 430 2.0 Array (Affymetrix).The Probe Array was scanned using a GeneChip(R) Scanner 3000 7G
2013-07-08 | E-GEOD-48595 | biostudies-arrayexpress
Project description:Cryptococcus gattii genome sequencing
| PRJNA660464 | ENA
Project description:In Trypanosoma brucei, there are fourteen enzymatic biotransformations that collectively convert glucose into five essential nucleotide sugars: UDP-Glc, UDP-Gal, UDP-GlcNAc, GDP-Man and GDP-Fuc. These biotransformations are catalyzed by thirteen discrete enzymes, five of which possess putative peroxisome targeting sequences. Published experimental analyses using immunofluorescence microscopy and/or digitonin latency and/or subcellular fractionation and/or organelle proteomics have localized eight and six of these enzymes to the glycosomes of bloodstream form and procyclic form T. brucei, respectively. Here we increase these glycosome localizations to eleven in both lifecycle stages while noting that one, phospho-N-acetylglucosamine mutase, also localizes to the cytoplasm. In the course of these studies, the heterogeneity of glycosome contents was also noted. These data suggest that, unlike other eukaryotes, all of nucleotide sugar biosynthesis in T. brucei is compartmentalized to the glycosomes in both lifecycle stages.
2021-01-28 | PXD023124 | Pride
Project description:Our previous investigation indicated that high-virulence C. gattii (C. gattii R265) tend to reside in the alveoli, whereas low-virulence C. gattii (C. gattii 5815) tend to be washed out from the alveoli and move into the central side of the respiratory system of the mice. Furthermore, C. gattii R265 and 5815 infected mice showed much lesser macrophage response than C. neoformans H99 infected mice. To elucidate the mechanism of this phenomenon from the viewpoint of genetic analysis, we performed this microarray assay. C. gattii R265, C. gattii 5815, and C. neoformans H99 were prepared for the study. The mice were anesthetized and the spore suspension was intratracheally injected into each mouse. Mice were sacrificed 15 days after the infection and then examined. Lungs of the mice infected with C. gattii R265 (n=4), C. gattii 5815 (n=4), and C. neoformans H99 (n=3) and of a normal control (n=1) were obtained. RNA targets preparation was performed according to the manufacturer's protocol using GeneChip(R) 3' IVT Express Kit (Affymetrix). One hundred nanograms of total RNA were converted into double-stranded cDNA template for transcription. In vitro transcription synthesized amplified RNA (aRNA) and incorporated a biotin-conjugated nucleotide. After purification and fragmentation of aRNA, 12.5 ug of them was hybridized to GeneChip(R) Mouse Genome 430 2.0 Array (Affymetrix).The Probe Array was scanned using a GeneChip(R) Scanner 3000 7G.
2016-06-22 | E-GEOD-83594 | biostudies-arrayexpress