Project description:We generated Arabidopsis lines expressing tagged (3X Myc tag) versions of BDR1 or BDR2 driven by their endogenous promoters on a bdrs triple mutant background (bdrs: cross of bdr1/AT5G25520 mutant SALK_142108C, bdr2/AT5G11430 mutant CS852350 and bdr3/AT2G25640 mutant SALK_059905). The genome-wide distribution of BDR1 and BDR2 in 8-day old seedlings were analyzed by ChIP-seq using an anti Myc antibody. We also analyzed by ChIP-seq the genome-wide distribution of FPA (AT2G43410) in wild-type seedlings using a rabbit anti-FPA polyclonal antibody raised against the C-terminal portion (position 536-901) of the FPA protein. Controls include sequencing of input DNA for each sample and a ChIP perfomed in wild-type seedlings with the anti-Myc tag antibody. We found that BDR1 and BDR2 are enriched at the border of a large number of genes in Arabidopsis genome and that FPA localizes mostly at the 3' end of genes.
Project description:We used an MNase digestion of chromatin from Arabidopsis seedlings, combined or not with ChIP (native ChIP), to analyze by high-throughput sequencing the genome-wide profiles of nucleosomes (MNase-seq), and of total H3, H3K4me2, H3K4me3 and H3K36me3 (native ChIPs) in wild-type (Col-0), fpa mutant (fpa/AT2G43410, line fpa-7) and a triple mutant of all three BDR proteins (bdrs: cross of bdr1/AT5G25520 mutant SALK_142108C, bdr2/AT5G11430 mutant CS852350 and bdr3/AT2G25640 mutant SALK_059905). We found that BDR proteins occupy regions of low nucleosome density. We also observed that genes upregulated in bdrs triple mutant display high levels of RNA polymerase II on their gene bodies but low levels of H3K4me3 and H3K36me3 in wild-type seedlings. For genes with the highest levels of BDR occupancy in wild-type, increased mRNA expression in bdrs mutant is associated with reduced RNA polymerase II density profile and increased H3K4me3 and H3K36me3 levels.
Project description:We used 3 different antibodies targeting the YSPTSPS repeats of the C-terminal domain (CTD) of the Rpb1 subunit of RNA polymerase II (ab817: non phosphorylated repeats, ab5095: phospho Ser2 repeats and ab5131: phospho Ser5 repeats) to obtain genome-wide profiles by ChIP-seq of the localization of RNA polymerase II in wild-type (Col-0), fpa mutant (fpa/AT2G43410, line fpa-7) and a triple mutant of all three BDR proteins (bdrs: cross of bdr1/AT5G25520 mutant SALK_142108C, bdr2/AT5G11430 mutant CS852350 and bdr3/AT2G25640 mutant SALK_059905). We found that BDR proteins are involved in RNA polymerase II 3' pausing and that their deletion generates transcriptional interferences in closely spaced, tandemly organized, gene pairs.
Project description:The aim of this experiment is to assess the genome-wide occupancy of Bye1, TFIIS and RNA polymerase II in yeast Saccharomyces cerevisiae by ChIP-chip