Project description:Dry skin evokes itch (pruritus), but underlying mechanisms are still elusive. To examine alterations of itch-related genes, dry skin condition was induced by AEW (Acetone/Ether/Water) treatment. In this study, RNA-seq was performed with dorsal root ganglia neurons obtained from AEW-treated ICR male mice (12-week-old) or control mice.
Project description:The large Drosophila protocadherin Fat (Ft) is a receptor for signal transduction pathways that control growth (Hippo signaling), planar cell polarity (PCP), metabolism and the proximodistal patterning of appendages. The intracellular domain (ICD) of Ft is crucial in implementing its biological functions. Six regions of high conservation (named A-F) within the ICD have been identified, as well as distinct regions mediating Hippo pathway activity that have been functionally characterized via overexpression assays. Here, we make targeted deletions of these highly conserved residues and the putative Hippo- and PCP-regulating domains of endogenous Ft using CRISPR/Cas9. Through transcriptomic, developmental, and phenotypic analyses, we show that different regions of Ft contribute uniquely to chromatin dynamics, tissue morphogenesis, PCP and metabolic regulation. We also demonstrate that different regions of Ft regulate Yorkie activity in opposite directions, finely tuning growth and tissue development. Strikingly, conserved regions D and F are key regulators of Ft’s functions- exhibiting opposing effects on wing morphology and Hippo pathway modulation- and conserved region D is the main regulator of PCP.
Project description:The large Drosophila protocadherin Fat (Ft) is a receptor for signal transduction pathways that control growth (Hippo signaling), planar cell polarity (PCP), metabolism and the proximodistal patterning of appendages. The intracellular domain (ICD) of Ft is crucial in implementing its biological functions. Six regions of high conservation (named A-F) within the ICD have been identified, as well as distinct regions mediating Hippo pathway activity that have been functionally characterized via overexpression assays. Here, we make targeted deletions of these highly conserved residues and the putative Hippo- and PCP-regulating domains of endogenous Ft using CRISPR/Cas9. Through transcriptomic, developmental, and phenotypic analyses, we show that different regions of Ft contribute uniquely to chromatin dynamics, tissue morphogenesis, PCP and metabolic regulation. We also demonstrate that different regions of Ft regulate Yorkie activity in opposite directions, finely tuning growth and tissue development. Strikingly, conserved regions D and F are key regulators of Ft’s functions- exhibiting opposing effects on wing morphology and Hippo pathway modulation- and conserved region D is the main regulator of PCP.
Project description:This series represents the analysis of a commercial dry active yeast (purchased locally). The genetics of this sample is unknown. Keywords = commercial Keywords = dry active yeast Keywords: other
Project description:The mutant Saccharomyces cerevisiae Y518 generated much more intracellular glutathione (GSH) than its counterpart dose. The RNA-seq based global transcriptome analysis was performed for exploring the potential mechanisms. Statistical analysis indicated that 1125 differentially expressed genes (fold-change>=2.0, FDR<=0.001) were up-regulated and 503 were down-regulated. There were 12 genes involved in glutathione metabolism. Of which GSH1, encodes gamma-glutamine cysteine synthetase, the rate-limiting enzyme in GSH biosynthesis process, was up-regulated. And MET17, encodes cysteine synthase A, an enzyme catalyzes the biosynthesis of one of the precursor amino acids L- cysteine, was also up-regulated. Besides, regulator SKN7 and several genes involved in oxidative stress response (GPX2, CTT1, SOD1, TRX2) were up-regulated. Intracellular ROS level of Y518 was also enhanced compared to that of 2-10515. Our results indicate that up-regulations of GSH1 and MET17 might be associated with the increased intracellular GSH content of the mutant, and up-regulated GSH1 may be caused by increased intracellular ROS level. Saccharomyces cerevisiae mRNA profiles of 24-h wild type 2-10515 and mutant Y518 were generated by deep sequencing using Illumina HiSeqTM 2000
Project description:Raw spectrum and search engine files to accompany JPR submitted manuscript entitled "Identification and Characterization of Human Proteoforms by Top-Down LC-21 Tesla FT-ICR Mass Spectrometry" by Anderson LC, DeHart CJ, Kaiser NK, Fellers RT, Smith DF, Greer JB, Blakney GT, Thomas PM, Kelleher NL, Hendrickson CL.
Project description:The 29-kDa RNA protein (29RNP), which was originally identified in rice etioplasts, has increased phosphorylation levels during de-etiolation. Here, using heparin-sepharose enriched fractions, we identified a chloroplast kinase that phosphorylated 29RNP. The chloroplast kinase phosphorylated the 29RNP and exhibited CKII biochemical characteristics in a kinase reaction test. Proteomic analysis of the chloroplast heparin-sepharose enriched fractions revealed the presence of 70 proteins, including both abundant proteins, such as plastid encoded polymerase subunits, and low-abundance proteins, such as APO2 and DAG. An inclusion list method using Fourier transform ion cyclotron resonance mass spectrometer ( FT-ICR LTQ MS ) was subsequently used to analyze the chloroplast heparin-sepharose enriched fractions. The 29RNP kinase was positively identified as a CKII alpha family protein by the presence of two unique peptides.