Project description:ChIP-chip profiles of JIL-1, H3S10phK14ac and H4K16ac in Drosophila S2 cells JIL-1, H3S10phK14ac and H4K16ac ChIP in Drosophila S2 cells. 2-4 biological replicates per experiment. dye-swaps as indicated in sample description
Project description:ChIP-chip profiles of RNA Polymerase II phosphorylated on serine 2 in Drosophila S2 cells. RNA Polymerase II phosphorylated on serine 2 ChIP in Drosophila S2 cells. 3 biological replicates with dye-swaps.
Project description:The Drosophila MSL complex mediates dosage compensation by increasing transcription of the single X chromosome in males approximately two-fold. This is accomplished through recognition of the X chromosome and subsequent acetylation of histone H4K16 on X-linked genes. Initial binding to the X is thought to occur at a subset of sites. However, the consensus sequence motif of entry sites (M-bM-^@M-^\MSL recognition elementM-bM-^@M-^] or MRE) is only slightly enriched on the X (~2 fold), and only a fraction of them is utilized by the MSL complex. Here we ask whether chromatin context could distinguish between utilized and non-utilized copies of the motif, by comparing their relative enrichment for histone modifications and chromosomal proteins mapped in the NHGRI modENCODE project. Through a comparative analysis of the chromatin features in male S2 cells, which contain MSL complex, and female Kc cells, which lack the complex, we find that the presence of active chromatin modifications, together with an elevated local GC content in surrounding sequence, has strong predictive value for functional MSL entry sites, independent of MSL binding. We tested these sites for function in Kc cells by RNAi knockdown of Sxl, resulting in induction of MSL complex. We show that ectopic MSL expression in Kc cells leads to H4K16 acetylation around these sites, and a relative increase in X chromosome transcription. Collectively, our results support a model in which a pre-existing active chromatin environment, coincident with H3K36me3, contributes to MSL entry site selection. The consequences of MSL targeting of the male X chromosome include increase in nucleosome lability, enrichment for H4K16 acetylation and JIL-1 kinase, and depletion of linker histone H1 on active X-linked genes. Our finding serves as a model to understand how chromatin and local sequence features are involved in the selection of functional protein binding sites in the genome. The key Drosophila female sex determinant protein, SXL, represses dosage compensation by inhibiting MSL2 translation. Loss of SXL results in the expression, stabilization, and targetting of the MSL complex in female cells. Therefore, depletion of SXL by RNA interference (RNAi) in female Kc cells will lead to a MSL2-dependent increase in transcription from the female X chromosomes, consistent with the induction of dosage compensation. In this experiment, we generated ChIP-chip profiles of H4K16 acetylation (H4K16ac) in Kc cells of control (GFP) and Sxl RNAi. For ChIP, we used Upstate (now Millipore) anti-H4K16ac antibody, catalog # 07-329, lot #JBC1355376.
Project description:modENCODE_submission_5619 This submission comes from a modENCODE project of Gary Karpen. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We aim to determine the locations of the major histone modifications across the Drosophila melanogaster genome. The modifications under study are involved in basic chromosomal functions such as DNA replication, gene expression, gene silencing, and inheritance. We will perform Chromatin ImmunoPrecipitation (ChIP) using genomic tiling arrays. We will initially assay localizations using chromatin from three cell lines and two embryonic stages, and will then extend the analysis of a subset of proteins to four additional animal tissues/stages. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf EXPERIMENT TYPE: CHIP-chip. BIOLOGICAL SOURCE: Strain: Sexed Female Jil mutant(official name : Jil-1 z2 genotype : Jil-1 z2/ Jil-1 z2 outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Developmental Stage: 3rd Instar Larvae Sexed Female ; Genotype: Jil-1 z2/ Jil-1 z2; Sex: Female; Transgene: deletion; NUMBER OF REPLICATES: 4; EXPERIMENTAL FACTORS: Strain Sexed Female Jil mutant(official name : Jil-1 z2 genotype : Jil-1 z2/ Jil-1 z2 outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Antibody H4K16ac(M) (target is H4K16ac); Developmental Stage 3rd Instar Larvae Sexed Female
Project description:modENCODE_submission_5620 This submission comes from a modENCODE project of Gary Karpen. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We aim to determine the locations of the major histone modifications across the Drosophila melanogaster genome. The modifications under study are involved in basic chromosomal functions such as DNA replication, gene expression, gene silencing, and inheritance. We will perform Chromatin ImmunoPrecipitation (ChIP) using genomic tiling arrays. We will initially assay localizations using chromatin from three cell lines and two embryonic stages, and will then extend the analysis of a subset of proteins to four additional animal tissues/stages. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf EXPERIMENT TYPE: CHIP-chip. BIOLOGICAL SOURCE: Strain: Sexed male Jil mutant(official name : Jil-1 z2 genotype : Jil-1 z2/Jil-1z2 outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Developmental Stage: 3rd Instar Larvae Sexed Male; Genotype: Jil-1 z2/Jil-1z2; Sex: Male; Transgene: deletion; NUMBER OF REPLICATES: 4; EXPERIMENTAL FACTORS: Strain Sexed male Jil mutant(official name : Jil-1 z2 genotype : Jil-1 z2/Jil-1z2 outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Antibody H4K16ac(M) (target is H4K16ac); Developmental Stage 3rd Instar Larvae Sexed Male
Project description:Profiling of changes in steady state RNA levels upon RNAi-mediated knockdown of the chromosomal kinase JIL-1 in Drosophila S2 cells.
Project description:modENCODE_submission_5622 This submission comes from a modENCODE project of Gary Karpen. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We aim to determine the locations of the major histone modifications across the Drosophila melanogaster genome. The modifications under study are involved in basic chromosomal functions such as DNA replication, gene expression, gene silencing, and inheritance. We will perform Chromatin ImmunoPrecipitation (ChIP) using genomic tiling arrays. We will initially assay localizations using chromatin from three cell lines and two embryonic stages, and will then extend the analysis of a subset of proteins to four additional animal tissues/stages. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf EXPERIMENT TYPE: CHIP-chip. BIOLOGICAL SOURCE: Strain: Sexed male Jil mutant-heterozygous(official name : JIL-1z2 genotype : Jil-1 z2/TM3 Ser-GFP outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Developmental Stage: 3rd Instar Larvae Sexed Male; Genotype: Jil-1 z2/TM3 Ser-GFP; Sex: Male; Transgene: deletion; NUMBER OF REPLICATES: 4; EXPERIMENTAL FACTORS: Strain Sexed male Jil mutant-heterozygous(official name : JIL-1z2 genotype : Jil-1 z2/TM3 Ser-GFP outcross : 1 transgene : deletion tags : Transposon tag : none description : - The JIL-1 allele JIL-1z2 was isolated in a screen for imprecise excisions of the EP transposon (Rørth et al. description : 1998) in w; EP(3)3657/?2?3 Sb heterozygotes Antibody H4K16ac(M) (target is H4K16ac); Developmental Stage 3rd Instar Larvae Sexed Male
Project description:modENCODE_submission_3038 This submission comes from a modENCODE project of Gary Karpen. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We aim to determine the locations of 125 chromosomal proteins across the Drosophila melanogaster genome. The proteins under study are involved in basic chromosomal functions such as DNA replication, gene expression, gene silencing, and inheritance. We will perform Chromatin ImmunoPrecipitation (ChIP) using genomic tiling arrays. We will initially assay localizations using chromatin from three cell lines and two embryonic stages, and will then extend the analysis of a subset of proteins to four additional animal tissues/stages For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Keywords: CHIP-chip EXPERIMENT TYPE: CHIP-chip. BIOLOGICAL SOURCE: Cell Line: S2-DRSC; Tissue: embryo-derived cell-line; Developmental Stage: late embryonic stage; Sex: Male; NUMBER OF REPLICATES: 4; EXPERIMENTAL FACTORS: Cell Line S2-DRSC; Antibody JIL-1(Q4170) (target is JIL-1)
Project description:Profiling of changes in steady state RNA levels upon RNAi-mediated knockdown of the chromosomal kinase JIL-1 in Drosophila S2 cells. Drosophila S2 cells were incubated 7 days after treatment with 10 µg of dsRNA directed against GST/EGFP or JIL-1, respectively. 5 biological replicates per target have been collected.