Transcription profiling of human galactose-1-phosphate uridyltransferase (GALT)-deficient cells reveals novel mechanisms of galactose toxicity
Ontology highlight
ABSTRACT: Comparison of gene expression profiles in GALT-deficient and GALT-reconstituted cells showed that cells lacking GALT activity responded to galactose challenge by activating a set of genes characteristic of endoplasmic reticulum (ER) stress. This response was specific to galactose insult, as cells grown in glucose or hexose-free media did not exhibit ER stress. Experiment Overall Design: Cells were first grown and then infected in high glucose, then transferred to DMEM, 3 experiments with dye-swap
Project description:Comparison of gene expression profiles in GALT-deficient and GALT-reconstituted cells showed that cells lacking GALT activity responded to galactose challenge by activating a set of genes characteristic of endoplasmic reticulum (ER) stress. This response was specific to galactose insult, as cells grown in glucose or hexose-free media did not exhibit ER stress. Keywords: GALT infected cells
Project description:To better understand the pathophysiology of galactose-1-phosphate uridyltransferase (GALT) deficiency in humans, we studied the mechanisms by which a GALT-deficient yeast survived on galactose medium. Under normal conditions, GALT-deficient yeast cannot grow in medium that contains 0.2% galactose as the sole carbohydrate, a phenotype of Gal(-). We isolated revertants from a GALT-deficient yeast by direct selection for growth in galactose, a phenotype of Gal(+). Comparison of gene expression profiles among wild-type and revertant strains on galactose medium revealed that the revertant down-regulated genes encoding enzymes including galactokinase, galactose permease, and UDP-galactose-4-epimerase (the GAL regulon). By contrast, the revertant strain up-regulated the gene for UDP-glucose pyrophosphorylase, UGP1. There was reduced accumulation of galactose-1-phosphate in the galactose-grown revertant cells when compared to the GALT-deficient parent cells.
Project description:To understand the altered gene expression by D-galactose in galT cells, the cells were grown in the presence and absence of D-galactose and tiling array analysis was performed for transcriptome profiling. A galT defect causes the disease Galactosemia in human. This work helps us to study the human metabolic disease Galactosemia. Total RNAs were isolated from Escherichia coli galT strain grown in the absence and presence of D-galactose. The transcriptome profile of galT cells was figured out in the galactose-stressed condition.
Project description:To understand the altered gene expression by D-galactose in galT cells, the cells were grown in the presence and absence of D-galactose and tiling array analysis was performed for transcriptome profiling. A galT defect causes the disease Galactosemia in human. This work helps us to study the human metabolic disease Galactosemia.
Project description:Comparison of the Streptococcus pneumoniae D39 wild type in M17 medium+ 0.5 % (w/v) Galactose (GaM17) compared to M17 medium+ 0.5 % (w/v) Glucose (GM17) Two condition design comparison of Wild-type strain including a dye swap
Project description:Xenotransplantation holds the promise of providing an unlimited supply of donor organs for terminal patients with organ failure. The gal carbohydrate results in rejection of wild type pig grafts, however, chimerism established by expression of the GalT gene prior to transplantation in GalT knockout mice results in tolerance to Gal+ heart grafts. We used microarrays in order to further understand the early events that occur within grafts that demonstrate tolerance. Experiment Overall Design: The GalT BMT recipient is a GalT knockout mouse which recieved GalT gene transduced allo-bone marrow cells transplantation after sublethal irradiation. A heart of wild type C57BL/6 was heterotopically transplanted into the recipient after GalT BMT. Syngeneic Control recipient is a wild type C57BL/6 transplanted a heart of wild C57BL/6.
Project description:Due to shortage of donor kidney, genetically modified kidney xenograft from pigs is considered. To select the optimal gene modification for xenotransplantation, knockout of carbohydrate genes or knockin of protective proteins have been applied. In this study, we utilized GalT- /-;hCD39;hCD55 and GalT-/-;hCD39;hCD46;hCD55;thrombomodulin (TBM) pigs for transplantation in non-human primates. NHPs survived for 4 weeks after kidney transplantation (4 WAT) from GalT-/-;hCD39;hCD55 pig and 6 WAT from GalT- /-;hCD39;hCD46;hCD55;TBM pig. However, mRNA sequencing and immunohistochemistry analysis revealed that 6 WAT kidney showed the severe apoptosis, inflammation, loss of renal function, and renal fibrosis than 4 WAT kidney. These results suggest that additional knockin of anti-complement (hCD26) and anti-coagulation (TBM) is not sufficient to inhibit the renal damage.
Project description:Transcriptome comparison of the Streptococcus pneumoniae D39 wild-type grown in M17 medium + 0.5 % (w/v) Galactose (GaM17) to grown in M17 medium + 0.5 % (w/v) Glucose (GM17).
Project description:This dataset contains single-cell RNA sequencing data from lung tissues of Galt gene-edited (GAL) mice and wild-type (WT) mice. The samples were collected for the investigation of molecular changes induced by Galt gene mutations, specifically focusing on lung injury. The dataset includes gene expression profiles of individual cells and enables the study of cell-type-specific responses, immune cell changes, and alterations in key cellular pathways. This dataset is useful for further understanding the pathophysiology of lung injury associated with Galt gene mutations and provides a resource for research into the molecular mechanisms of galactosemia.
Project description:We report RNAseq data from B. salamandrivorans zoospores treated with different carbohydrates (glucose, mannose or galactose, 50 mM) or H2O as a control, for 1 hour. We found a selective upregulation of putative B. salamandrivorans virulence factors during initial contact with carbohydrates and galactose specifically.