Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

H3K36 mutations promote sarcomagenesis through genome-wide remodeling of H3K36 and H3K27 methylation

ABSTRACT: This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Peter Lewis

PROVIDER: E-GEOD-69291 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Histone H3K36 mutations promote sarcomagenesis through altered histone methylation landscape.

Lu Chao C Jain Siddhant U SU Hoelper Dominik D Bechet Denise D Molden Rosalynn C RC Ran Leili L Murphy Devan D Venneti Sriram S Hameed Meera M Pawel Bruce R BR Wunder Jay S JS Dickson Brendan C BC Lundgren Stefan M SM Jani Krupa S KS De Jay Nicolas N Papillon-Cavanagh Simon S Andrulis Irene L IL Sawyer Sarah L SL Grynspan David D Turcotte Robert E RE Nadaf Javad J Fahiminiyah Somayyeh S Muir Tom W TW Majewski Jacek J Thompson Craig B CB Chi Ping P Garcia Benjamin A BA Allis C David CD Jabado Nada N Lewis Peter W PW

Science (New York, N.Y.) 20160501 6287

Several types of pediatric cancers reportedly contain high-frequency missense mutations in histone H3, yet the underlying oncogenic mechanism remains poorly characterized. Here we report that the H3 lysine 36-to-methionine (H3K36M) mutation impairs the differentiation of mesenchymal progenitor cells and generates undifferentiated sarcoma in vivo. H3K36M mutant nucleosomes inhibit the enzymatic activities of several H3K36 methyltransferases. Depleting H3K36 methyltransferases, or expressing an H3 ...[more]

PMID: 27174990

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Project description:Identification and characterization of HP1BP3 (a human histone H1 homologue) as a novel chromatin retention factor essential for the co-transcriptional processing of pri-miRNA. We generated BAC transgenic cells at 80% confluency (~1x107) were cross-linked with 1% formaldehyde for 10 minutes at 37Â°C, and quenched with 125 mM glycine at room temperature for 5 minutes. The fixed cells were washed twice with cold PBS, scraped, and transferred into 1 ml PBS containing protease inhibitors (Roche). After centrifugation at 700 g for 4 minutes at 4Â°C, the cell pellets were resuspended in 100 Î¼l ChIP lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl [pH 8.1] with protease inhibitors) and sonicated at 4Â°C with a Bioruptor (Diagenode) (30 seconds ON and 30 seconds OFF at highest power for 15 minutes). The sheared chromatin with a fragment length of ~200 â 600 bp) was centrifuged at 20,000 g for 15 minutes at 4Â°C). 100 Î¼l of the supernatant was used for ChIP or as input. A 1:10 dilution of the solubilized chromatin in ChIP dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 167 mM NaCl 16.7 mM Tris-HCl [pH 8.1]) was incubated at 4Â°C overnight with 6 Î¼g/ml of a goat anti-GFP (raised against His-tagged full-length eGFP and affinity-purified with GST-tagged full-length eGFP). Immunoprecipitation was carried out by incubating with 40 Î¼l pre-cleared Protein G Sepharose beads (Amersham Bioscience) for 1 hour at 4Â°C, followed by five washes for 10 minutes with 1ml of the following buffers: Buffer I: 0.1% SDS, 1% Triton X-100, 2 mM EDTA, 20 mM Tris-HCl [pH 8.1], 150 mM NaCl; Buffer II: 0.1% SDS, 1% Triton X-100, 2 mM EDTA, 20 mM Tris-HCl [pH 8.1], 500 mM NaCl; Buffer III: 0.25 M LiCl, 1% NP-40, 1% deoxycholate, 1 mM EDTA, 10 mM Tris-HCl [pH 8.1]; twice with TE buffer [pH 8.0]. Elution from the beads was performed twice with 100 Î¼l ChIP elution buffer (1% SDS, 0.1 M NaHCO3) at room temperature (RT) for 15 minutes. Protein-DNA complexes were de-crosslinked by heating at 65Â°C in 192 mM NaCl for 16 hours. DNA fragments were purified using QiaQuick PCR Purification kit (QIAGEN) and eluted into 30 Î¼l H2O according to the manufacturerâs protocol after treatment with RNase A and Proteinase K.

Dataset Information

H3K36 mutations promote sarcomagenesis through genome-wide remodeling of H3K36 and H3K27 methylation

Publications

Histone H3K36 mutations promote sarcomagenesis through altered histone methylation landscape.

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