RNAseq of Arabidopsis plants lacking E2 ubiquitin conjugating enzymes UBC35 and UBC36
Ontology highlight
ABSTRACT: This RNAseq experiment aims at determining the reprogramming of gene expression upon loss of K63 polyubiquitin chain formation. Twelve-day-old plants expressing a beta-estradiol inducible artificial microRNA targeting the two plant E2 enzymes catalyzing K63 polyubiquitin chain formation were mock-treated or beta-estradiol-treated (5microM). Total RNAs from whole seedings were extracted and subjected to RNA-seq analyses. Genes that are differentially regulated between non-treated (uninduced) and treated (induced with beta-estradiol) were identified.
Project description:RP-LC-MS lipidomics data was collected to understand the role of GATA1 during erythroid maturation. GATA1 mutants and WT cells were treated with or without beta-estradiol. GATA1 mutant cells were additionally treated with or without 5-ALA.
Project description:RP-LC-MS lipidomics data was collected to understand the role of GATA1 during erythroid maturation. GATA1 mutants and WT cells were treated with or without beta-estradiol. GATA1 mutant cells were additionally treated with or without 5-ALA.
Project description:Total RNA was analyzed from either uninduced or β-estradiol treated G1E-ER-GATA cells to determine changes in gene expression upon induction of erythroid maturation (treated).
Project description:Alas2 gene encodes the rate-limiting enzyme in heme biosynthesis. CRISPR/Cas9-mediated ablation of two Alas2 intronic cis-elements strongly reduced GATA-1-induced Alas2 transcription, heme biosynthesis, and GATA-1 regulation of other vital constituents of the erythroid cell transcriptome. Bypassing Alas2 function in Alas2 cis-element-mutant (double mutant) cells by providing its catalytic product 5-aminolevulinic acid (5-ALA) rescued heme biosynthesis and the GATA-1-dependent genetic network. We discovered a GATA factor- and heme-dependent circuit that establishes the erythroid cell transcriptome. G1E-ER-GATA-1 WT and double mutant cells were examined. Untreated WT, beta-estradiol-treated WT, beta-estradiol-treated double-mutant, and beta-estradiol/5-ALA-treated double-mutant cells were subjected to RNA-seq.
Project description:A polyubiquitin chain can adopt a variety of shapes, depending on how the ubiquitin monomers are joined. However, the relevance of linkage for the signaling functions of polyubiquitin chains is often poorly understood because of our inability to control or manipulate this parameter in vivo. Here we present a strategy for reprogramming polyubiquitin chain linkage by means of tailor-made, linkage- and substrate-selective ubiquitin ligases. Using the polyubiquitylation of the budding yeast replication factor PCNA in response to DNA damage as a model case, we show that altering the features of a polyubiquitin chain in vivo canchange the fate of the modified substrate. We also provide evidence for redundancy betweendistinct, but structurally similar linkages, and we demonstrate by proof-of-principle experiments that the method can be generalized to targets beyond PCNA. Our study illustrates apromising approach towards the in vivo analysis of polyubiquitin signaling.
Project description:The purpose of this study was to characterize the transcriptional effects induced by intramuscular IFN-beta-1a treatment (Avonex, 30 µg once weekly) in patients with relapsing-remitting form of multiple sclerosis (MS). By using Affymetrix DNA microarrays, we obtained genome-wide expression profiles of peripheral blood mononuclear cells from 24 MS patients within the first four weeks of IFN-beta administration. Keywords: Multiple sclerosis, Interferon, Pharmacogenomics, Affymetrix EDTA blood samples were taken from all patients immediately before first, second and fifth IFN-beta injection. Total RNA of Ficoll-isolated peripheral blood mononuclear cells from each sample was extracted, labelled and hybridized to Affymetrix Human Genome U133 A and B arrays to quantify the mRNA levels.
Project description:Publication upload for journal submission 'The Ubiquitin Interacting Motif Like Domain of Met4 is a K48 Polyubiquitin Chain Selective Binding Domain'
Project description:We defined the C/EBPa signature characterized by a set of genes which are upregulated upon C/EBPa activation. In order to identify the C/EBPa signature, we performed microarray gene expression analysis of K562 cells stably expressing p42-C/EBPa-ER after activating the C/EBPa construct to translocate to the nucleus for 6 hours with beta-estradiol. The gene expression profile was performed in K562 p42-C/EBPa-ER expressing cells treated with beta-estradiol (n=4) or EtOH vehicle control (n-4) for 6 hours. Induction of nuclear localization of C/EBPa-ER fusion protein was achieved by addition of 1 uM beta-estradiol as indicated into the culture medium.