Project description:Transcriptional profiling of 16-day-old seedlings of Arabidopsis wild type control and mutants is performed using AligentM-bM-^@M-^Ys Whole Arabidopsis Gene Expression Microarray (4x44K). Two-condition experiment, seedlings of wild type control vs. Mutant sdg25, sdg26, sdg25 sdg26, atx1, sdg26 atx1, clf, sdg26 clf, ldl1 ldl2, sdg25 ldl1 ldl2 or sdg26 ldl1 ldl2. Three biological replicates: 3 control, 3 each of the ten mutants, independently grown under 12h light/ 12h dark photoperiods and harvested.
Project description:This SuperSeries is composed of the following subset Series: GSE24646: h3k36me3-Establishing a reference epigenome in arabidopsis seedlings GSE24647: h3k9me2-Establishing a reference epigenome in arabidopsis seedlings GSE24648: h3k4me2-Establishing a reference epigenome in arabidopsis seedlings GSE24649: h3k4me3-Establishing a reference epigenome in arabidopsis seedlings GSE24650: h3k9me3-Establishing a reference epigenome in arabidopsis seedlings GSE24651: h3k27me3-Establishing a reference epigenome in arabidopsis seedlings GSE24652: h3k27me2-Establishing a reference epigenome in arabidopsis seedlings GSE24653: h3k56ac-Establishing a reference epigenome in arabidopsis seedlings GSE24654: h4k20me1-Establishing a reference epigenome in arabidopsis seedlings GSE24655: h3-Establishing a reference epigenome in arabidopsis seedlings GSE24656: h2bub-Establishing a reference epigenome in arabidopsis seedlings GSE24657: h3k27me3_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24658: h3k4me3_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24659: h3k4me2_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24660: 5mc_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24661: h3k36me3_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24662: h3k27me1_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24663: h3_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24664: h3k27me3_roots_gw-Establishing a reference epigenome in arabidopsis seedlings GSE24665: h3k4me3_roots_gw-Establishing a reference epigenome in arabidopsis seedlings Refer to individual Series
Project description:Polycomb Group Proteins (PcGs) is critical in defining the epigeneticÂ blueprintÂ for animal and plant development. In plants, loss of different PcGs display both common and unique phenotypic defects, yet little is known about how these are established. Here, based on quantitative comparison of epigenomics data from mutants of key PcG components in Arabidopsis seedlings, we found that the PcG partners of CURLY LEAF (CLF), one of the major plant H3K27 trimethyltransferases, determines its selectivity in repressing gene loci involved in distinct developmental programs. The non-redundant role of CLF in determining flower development is specifically associated with HETEROCHROMATIN PROTEIN1 (LHP1). This context dependent effect of CLF corresponds well with tissue-biased target gene expression, and importantly, to differential co-occupancy of transcription factors, such as MADS box and B3-domain transcription factors. These results provide valuable insight as to the dynamic interplay between different PcGs and their collaborative control of plant development. To compare the effect of different PcGs on epigenetic structure from the genome-wide scale, we used chromatin immunoprecipitation followed by high-throughtput sequencing (ChIP-seq) to characterize the genome-wide binding profile of H3K27me3 in Col, clf-29, tfl2-2, atbmi1a/b and atring1a/b ; To investigate the functional consequence of the distinct H3K27me3 profile controlled by different combinations of PcGs, we characterized the transcriptome change in PcG mutants, including Col, clf-29, tfl2-2, lhp1-6, atbmi1a/b, atring1a/b, and clf29swn21.
Project description:Chromatin remodeling factors of the Imitation Switch (ISWI) family play important roles in epigenetic regulations of gene expression in yeast and animals, whereas their function in plants remains elusive. Here we report characterization of the Arabidopsis ISWI genes CHR11 and CHR17. Double mutant chr11 chr17 displayed a dramatically reduced plant size with early flowering. In addition, epidermis of the double mutant leaves showed cell characteristics seen only in floral organs. These phenotypes resemble, at least partially, those of the Polycomb mutants curly leaf (clf) and like heterochromatin protein1 (lhp1). Microarray analysis revealed that a number of targets of the Polycomb pathway were derepressed in chr11 chr17 leaves. Furthermore, triple mutants combining chr11 chr17 with clf-29 or lhp1-6 both greatly enhanced clf-29 and lhp1-6 phenotypes, respectively. All these results strongly suggest that the ISWI family genes in Arabidopsis may function in gene silencing via the Polycomb pathway Leaves from 15-day-old seedlings of wild-type Col-0 and chr11-1 chr17-1 were used for RNA preparation.In the experiment data,A refers to Col-0, and B refers to chr11-1 chr17-1.
Project description:The Polycomb Group (PcG) proteins form two protein complexes, PcG Repressive Complex 1 (PRC1) and PRC2, which are key epigenetic regulators in eukaryotes. PRC2 represses gene expression by catalyzing the trimethylation of histone H3 lysine 27 (H3K27me3). In Arabidopsis (Arabidopsis thaliana), CURLY LEAF (CLF) and SWINGER (SWN) are two major H3K27 methyltransferases and core components of PRC2, playing essential roles in plant growth and development. Despite their importance, genome-wide binding profiles of CLF and SWN have not been determined and compared yet. In this study, we generated transgenic lines expressing GFP-tagged CLF/SWN under their respective native promoters and used them for ChIP-seq analyses to profile the genome-wide distributions of CLF and SWN in Arabidopsis seedlings. We also profiled and compared the global H3K27me3 levels in wild-type (WT) and PcG mutants (clf, swn, and clf swn). Our data show that CLF and SWN bind to almost the same set of genes, except that SWN has a few hundred more targets. Two short DNA sequences, the GAGA-like and Telo-box-like motifs, were found enriched in the CLF and SWN binding regions. The H3K27me3 levels in clf, but not in swn, were markedly reduced compared with WT; and the mark was undetectable in the clf swn double mutant. Further, we profiled the transcriptomes in clf, swn, and clf swn, and compared that with WT. Thus this work provides a useful resource for the plant epigenetics community for dissecting the functions of PRC2 in plant growth and development.
Project description:CURLY LEAF (CLF), the major histone methyltransferase of Polycomb Repressive Complex 2 (PRC2), modifies trimethylation of histone H3 lysine 27 (H3K27me3) and mediates dynamical chromatin repression in Arabidopsis. Here we used strand specific RNA-sequencing to profile Arabidopsis transcriptomes obtained from roots, shoots, flowers and siliques of Col-0 and clf-28 plants. Our analysis identified a large number of CLF-regulatedd transcripts in Arabidopsis. Transcriptome profiling in roots, shoots, inflorescences and siliques of WT and clf-28 plants with 3 biological replicates.
Project description:Part of a set of highly integrated epigenome maps for Arabidopsis thaliana. Keywords: Illumina high-throughput transcriptome sequencing Transcriptome sequencing of wildtype Arabidopsis plants (Columbia-0), and met1, drm1 drm2 cmt3, and ros1 dml2 dml3 null mutants using the Illumina Genetic Analyzer.
Project description:Transcriptional profiling of 16-day-old seedlings of Arabidopsis wild type control and mutants is performed using Aligent’s Whole Arabidopsis Gene Expression Microarray (4x44K). Two-condition experiment, seedlings of wild type control vs. Mutant sdg25, sdg26, sdg25 sdg26, atx1, sdg26 atx1, clf, sdg26 clf, ldl1 ldl2, sdg25 ldl1 ldl2 or sdg26 ldl1 ldl2. Three biological replicates: 3 control, 3 each of the ten mutants, independently grown under 12h light/ 12h dark photoperiods and harvested.
Project description:Polycomb repressive complexes (PRCs) play crucial roles in transcriptional repression and developmental regulation in both plants and animals. In plants, depletion of different members of PRCs causes both overlapping and unique phenotypic defects. However, the underlying molecular mechanism determining the target specificity and functional diversity is not sufficiently characterized. Here, we quantitatively compared changes of tri-methylation at H3K27 in Arabidopsis mutants deprived of various key PRC components. We show that CURLY LEAF (CLF), a major catalytic subunit of PRC2, coordinates with different members of PRC1 in suppression of distinct plant developmental programs. We found that expression of flower development genes is repressed in seedlings preferentially via non-redundant role of CLF, which specifically associated with LIKE HETEROCHROMATIN PROTEIN1 (LHP1). In contrast, expression of embryo development genes is repressed by PRC1-catalytic core subunits AtBMI1 and AtRING1 in common with PRC2-catalytic enzymes CLF or SWINGER (SWN). This context-dependent role of CLF corresponds well with the change in H3K27me3 profiles, and is remarkably associated with differential co-occupancy of binding motifs of transcription factors (TFs), including MADS box and ABA-related factors. We propose that different combinations of PRC members distinctively regulate different developmental programs, and their target specificity is modulated by specific TFs.
Project description:We have identified a histone methyltransferase gene CURLY LEAF (CLF), which is required to repress a floral homeotic gene AGAMOUS (AG). CLF acts redundantly with a related gene SWINGER (SWN), so that clf swn double mutants have extreme, near lethal phenotypes in which expression of many target genes is disrupted. To understand better when and where these genes act, we propose to use a system for generating mosaic plants in which marked sectors of cells are created that gain or lose CLF (or other Pc-G) gene function in SWN+ and swn- mutant backgrounds. This will allow us to address questions such as whether the Pc-G genes are needed throughout development to silence their targets, whether they are needed once cell division has ceased, whether they can silence their targets once the targets have been activated, and whether genes that are epigenetically silenced by CLF are impervious to activating factors. Aim is to compare gene expression in wild-type and polycomb mutant seedlings. Comparison of bulked seedlings of four genotypes: wild-type (Columbia), clf-28, swn-7, clf-28 swn-7 double mutant. 8 samples were used in this experiment, 2 of each genotype. Tissue grown in tissue culture.