Project description:With the evolution of the PylRS system, we identified the PylRS variants for incorporation of all synthesized Kacyl and Kacyl* in the full-length H3 protein both in E. coli and mammalian cells, which covered the tail domain (eg. K23, K27, K56) as well as the globular domain that are difficult for chemical synthesis (eg. K64, K79, K122).
Project description:ChIP-chip assays to determine the occupancy of acetylated histone H3 K56 in wildtype, isw1, chd1, isw1 chd1 and ISW1[K227R] yeast. Two color experiment. ChIP/Input. K56ac data normalized to histone H3 and plotted as mutant over wildtype. Biological replicates=3. Please note that H3 data for YMS123-125 & YMS127 is part of the AcH4 file.
Project description:Purpose - To establish the response of B. cenocepacia K56-2 to growth in copper, and to characterise the BCAM0442/3 copper-sensing TCS. Methods - RNA-seq of the response WT and Δbcam0442/3 B. cenocepacia K56-2 to mid-log phase growth in LB with or without 1 mM CuCl2 was performed. Results - Characterisation of the overall response of B. cenocepacia K56-2 to copper, as well as establishing that BCAM0442/3 regulates the CopABCDE system in B. cenocepacia
Project description:Eukaryotic genomes are repetitively packaged into chromatin by nucleosomes, however they are regulated by the differences between nucleosomes, which establish various chromatin states. Replication-independent histone exchange could potentially perturb chromatin status if histone exchange chaperones, such as Swr1C, loaded histone variants into wrong sites. Here we use ChIP-chip analysis of H2A.Z-myc to show that in S.pombe, like S.cerevisiae, Swr1C is required for loading H2A.Z into specific sites, including the promoters of lowly expressed genes. However S.pombe Swr1C has an extra subunit, Msc1, which is a JumonjiC-domain protein of the Lid/Jarid1 family. The absence of Msc1 does not disrupt the S.pombe Swr1C or H2A.Z distribution in euchromatin. However in the absence of either Msc1 or Swr1 H2A.Z is ectopically found in the inner centromere and in subtelomeric chromatin. Normally this subtelomeric region, which appears to be a novel chromatin domain that we term ST-chromatin, not only lacks H2A.Z but also shows uniformly lower levels of H3K4me2, H4K5 and K12K5 acetylation than euchromatin. It also disproportionately contains the most lowly expressed genes during vegetative growth, including many meiotic-specific genes. These data describe H2A.Z distribution in S.pombe and identify a new mode of chromatin surveillance and maintenance based on negative regulation of histone variant misincorporation.
Project description:ChIP-on chip assays to measure the change in histone H3 K56 acetylation over the yeast genome in wild-type YBL574 yeast strains compared to H3K36A mutant strains. Two color experiment. Mutant vs WT cells. Biological replicates=3 per IP per cell type.