Project description:The quantitative proteomes of leaf tissue from A. thaliana at 4 weeks and 5 weeks of growth using tandem mass spectrometry with label-free quantification. A comparison of three genotypes has been performed, namely i) the Col-0 wild type ecotype, ii) a T-DNA insertion mutant disrupting the plastoglobule-localized ABC1K6 protein (abc1k6-1) that is delayed in transition to reproductive growth, and iii) a complementation line (Comp3) that restores the timely progression of development with the ABC1K6 wild-type protein sequence tagged C-terminally with GFP in the abc1k6-1 background. Whereas wild-type and abc1k6-1 remain in the vegetative growth stage at 4 weeks of age, the wild-type plants have transitioned to reproductive growth (as evidenced by the emergence of the infloresence bolt) by 5 weeks of age, while abc1k6-1 remians in vegetative growth. Our proteomics experiment seeks to identify leaf proteome differences prior to wild-type transition to reproductive growth that may account for the delayed transition in the abc1k6-1 mutant.
Project description:Total RNA was isolated with TRIZOL reagent from wild-type flies (yellow white, yw) and Pumilio mutant flies (pum13; Barker DD. et al. (1992) Genes Dev 6, 2312-26). 12 microgram of total RNA was used for cDNA synthesis using a 1:1 mixture of oligo(dT) and random nonamer primers and in the presence of amino-allyl dUTP. Set of arrays that are part of repeated experiments Keywords: Biological Replicate
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.
