Project description:Ascorbic acid has been reported to stimulate DNA iterative oxidase TET enzymes, Jumonji C-domain-containinghistone demethylase and potentially RNA m6A demethylase FTO and ALKBH5 as a cofactor. Although ascorbic acid has been widely investigated in reprogramming DNA and histone methylation status in vitro, in cell lines and mouse models, its specific role in the catalytic cycle of dioxygenases remains enigmatic. Here we systematically investigated the stimulation of ascorbic towards TET2, ALKBH3, histone demethylases and FTO. We find that ascorbic acid reprograms epitranscrip-tome by erasing the hypermethylated m6A sites. Biochemistry and Electron spin resonance (ESR) assays demonstrate that ascorbic acid enters the active pocket of dioxygenases, reduces Fe (III), either incorporated upon protein synthesis or generated upon rebounding the hydroxyl radical during oxidation, into Fe (II). Finally, we propose a new model for the catalytic cycle of dioxygenases by adding in the essential dynamic cofactor, ascorbic acid. Ascorbic acid refreshes and regenerates inactive dioxygenase through recycling Fe (III) into Fe (II) in a dynamic “hit-and-run” manner.
Project description:Ascorbic acid has been reported to stimulate DNA iterative oxidase TET enzymes, Jumonji C-domain-containinghistone demethylase and potentially RNA m6A demethylase FTO and ALKBH5 as a cofactor. Although ascorbic acid has been widely investigated in reprogramming DNA and histone methylation status in vitro, in cell lines and mouse models, its specific role in the catalytic cycle of dioxygenases remains enigmatic. Here we systematically investigated the stimulation of ascorbic towards TET2, ALKBH3, histone demethylases and FTO. We find that ascorbic acid reprograms epitranscrip-tome by erasing the hypermethylated m6A sites. Biochemistry and Electron spin resonance (ESR) assays demonstrate that ascorbic acid enters the active pocket of dioxygenases, reduces Fe (III), either incorporated upon protein synthesis or generated upon rebounding the hydroxyl radical during oxidation, into Fe (II). Finally, we propose a new model for the catalytic cycle of dioxygenases by adding in the essential dynamic cofactor, ascorbic acid. Ascorbic acid refreshes and regenerates inactive dioxygenase through recycling Fe (III) into Fe (II) in a dynamic “hit-and-run” manner.
Project description:Lactic acid is used to check skin sensitivity of human. However transcriptional profiling has not been done yet. This study was targeted to understand the transcriptional changes occur in human skin upon treatment with lactic acid.
Project description:Subgroup Ib BHLH genes are induced by Fe deficiency. BHLH038 and BHLH039 were shown to be connected with salicylic acid. The triple knockout 3xbhlh (bhlh039-1 bhlh100-1 bhlh101-1) shows a stronger leaf chlorosis at - Fe than the wild type. The responses of this triple mutant were studied at - Fe in comparison to the wild type in the presence and absence of salicylic acid (SA).
Project description:An isogenic canola line was repeatedly treated with Salicylic Acid and the most vital plants were selected for propagation. 2 SA resistant lines were produced. ChIP-Seq was performed on those two lines and the mother line 6 hours after SA treatment to check for different reactions. Antibodies against H3K4me3 and Pol2 were used to check for changes in epigenetic makeup and transcriptional response.
Project description:Subgroup Ib BHLH genes are induced by Fe deficiency. BHLH038 and BHLH039 were shown to be connected with salicylic acid. The triple knockout 3xbhlh (bhlh039-1 bhlh100-1 bhlh101-1) shows a stronger leaf chlorosis at - Fe than the wild type. The responses of this triple mutant were studied at - Fe in comparison to the wild type in the presence and absence of salicylic acid (SA). Wild type and 3xbhlh seedlings (bhlh039-1 bhlh100-1 bhlh101-1) were grown at M-bM-^@M-^S Fe and treated for six hours with or without 100 M-BM-5M SA. Three biological replicates were generated.