Project description:Transcriptome of seeds at eight days after polinization were compared between wild type and loss-of-function mutant in the flavonoid 3' hydroxylase TT7 gene
Project description:Sterol 27-hydroxylase (Cyp27a1) is a primary enzyme that is involved in the Alternative pathway of bile acid synthesis, which is particularly enhanced during pregnancy. However, the role of this enzyme during fetal organ formation was unclear. Here, we demonstrate that 27-hydroxylase activity in the mother is essential for the progression of pregnancy and organ formation in the fetuses. Maternal depletion of Cyp27a1 reduced success pregnancy rate and litter size. In addition, all of the delivered newborn mice died due to respiratory distress syndrome resulting from the absence of mature alveolar epithelial cells. These phenotypes were caused by 7α-hydroxycholesterol (7α-HC) accumulating in Cyp27a1 deficient mice. Mechanistically, 7α-HC bound to and destabilized a protein Fau, mediating the assembly of ribosomes, the down-regulation of which led to lower protein synthesis and polysome formation. Our study discovered the essential metabolic pathway protecting the developing fetuses from a toxic metabolite.
Project description:Collagen from the skin of wild type and prolyl 4-hydroxylase mutant mice was extracted and proline hydroxylation of the collagen-derived peptides were analyzed by LC-MS/MS.
Project description:Skeletal muscle has an impressive ability to repair itself after a damaging insult and this response is essential to the process of muscle adaptation. In conditions such as muscular dystrophy and the sarcopenia of old age, repair is compromised leading to fibrosis and fatty tissue accumulation. Hypoxia-inducible factors (HIFs) are highly conserved regulators of gene transcription under conditions of low oxygen tension and HIF target genes such as EPO and VEGF have been associated with muscle protection and repair. We sought to interrogate the importance of HIF activation to skeletal muscle repair through the use of prolyl hydroxylase inhibitors (PHI) that stabilize HIF and activate target gene transcription in a mouse eccentric exercise limb damage model. We used microarrays to detail the global effects of prolyl hydroxylase inhibitors (PHI) in a mouse eccentric exercise limb damage model.
Project description:To evaluate gene expression changes in response to prolyl hydroxylase domain protein 2 and 3 (PHD2/3) depletion in endothelial cells, we performed mRNA-sequencing analysis with heart samples isolated from PHD2/3 endothelial knockout mice and their littermate control wild type mice.
Project description:We performed shotgun proteomic analysis of type V collagens purified from wild-type mice and prolyl 3-hydroxylase 3 or lysyl hydroxylase 1 null mice by LC-MS after trypsin digestion for identification of lysine glycosylation sites.
Project description:Tyrosine hydroxylase (TH) is a highly regulated enzyme that catalyses the rate-limiting step in the biosynthesis of dopamine (DA) and other catecholamines. Mutations and dysfunction in this enzyme lead to DA deficiency and parkinsonisms of different severity. An understanding of TH deficiency at the level of structure and stability has been lacking to date, as only structures of truncated TH forms have been available. Here, we used cryoEM and XL-MS to determine the high-resolution structure of full-length human tetrameric TH in the absence and presence of the end-product and feedback inhibitor DA bound to the active site
Project description:Lysyl hydroxylase 2 (LH2) regulates the intermolecular cross-linking of collagen molecules. Accumulation of LH2-modified collagen, which is highly stable and resistant to collagenase cleavage, is one of the causes of fibrosis. In this study, we established LH2-deficient fibroblasts and first analyzed morphology of LH2-deficient fibroblasts by microscopy. LH2-deficient fibroblasts showed a dramatic increase in filopodial dynamics and cell migration, which were supported by time-lapse video microscopy and Immunocytochemistry, and gene expression profiling data. Understanding the precise role of phenotype in LH2-deficient cells may be helpful to define the pathogenesis of fibrosis.
