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Expression data from Suv420h1 knockout and mutant PREX2 expressing MEFs (mouse embryonic fibroblasts)

ABSTRACT: PREX2 truncating mutations occur in melanoma. We used microarray based gene expression profiling to compare expression patterns between cells harboring Suv420h1 knockout and PREX2 mutant expressing

ORGANISM(S): Mus musculus

PROVIDER: GSE57310 | GEO | 2016/04/06

SECONDARY ACCESSION(S): PRJNA246227

REPOSITORIES: GEO

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Expression data from Suv420h1 knockout and mutant PREX2 expressing MEFs (mouse embryonic fibroblasts)

Project description:PREX2 truncating mutations occur in melanoma. We used microarray based gene expression profiling to compare expression patterns between cells harboring Suv420h1 knockout and PREX2 mutant expressing Spontaneously immortalized MEFs from either wt or Suv420h1 knockout mice were used. Wt MEFs were transduced with lentivirus to express indicated PREX2 mutants. Suv420h1 ko MEFs were reconstituted either with control GFP or Suv420h1-GFP fusion. Total RNA was isolated and subjected to analysis

2016-04-06 | E-GEOD-57310 | biostudies-arrayexpress

Expression data from melanoma xenograft tumors originating from primary immortalized melanocytes

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Project description:PREX2 truncating mutations occur in melanoma. We used microarray based gene expression profiling to compare expression patterns between xenografts harboring control GFP, wt PREX2 or various human relevant PREX2 mutants Primary xenograft tumors derived from primary immortalized melanocytes expressing indicated constructs were grown in ncr-nude mice and tumors harvested before before reaching 1.5cm size

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A targetable PREX2/RAC1/PI3Kb signalling axis confers resistance to clinically relevant therapeutic approaches in melanoma

Project description:Metastatic melanoma remains a major clinical challenge. Large-scale genomic sequencing of melanoma has identified bona fide activating mutations in RAC1, with mutations of its upstream regulator, the RAC-GEF PREX2, also commonly detected. Crucially, RAC1 mutations are associated with resistance to BRAF-targeting therapies. Despite the role of its homologue PREX1 in melanomagenesis, and evidence that some truncating PREX2 mutations drive increased RAC1 activity, no hotspot mutations have been identified, and the impact of PREX2 mutation remains contentious. Here, we use genetically engineered mouse models and patient-derived BRAFV600E-driven melanoma cell lines to dissect the role of PREX2 in melanomagenesis and response to therapy. We show that while PREX2 is dispensable for the initiation and progression of melanoma, its loss confers sensitivity to clinically relevant therapeutics. Importantly, genetic and pharmacological targeting of the RAC1 effector kinase PI3Kβ phenocopies PREX2 loss, sensitizing our model systems to therapy. Our data reveal a druggable PREX2/RAC1/PI3Kβ signalling axis in BRAF-mutant melanoma that could be exploited clinically.

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Structural basis of nucleosomal H4K20 recognition and methylation by SUV420H1 methyltransferase

Project description:Here we report the cryo-electron microscopy (cryo-EM) structures of SUV420H1 associated with canonical nucleosome core particles (NCPs) or H2AZ containing NCPs. We find that SUV420H1 shows conformational change after bound to a nucleosome and make extensive site-specific contacts with histone and DNA regions, thus enabling H4K20 insertion for catalysis specifically. Through in vivo functional analysis, we validated special residues of SUV420H1 that are critical for its catalytic activity and function in chromatin state regulation.Thus, our study provides molecular insights into the nucleosome-based recognition and histone- methylation mechanisms of SUV420H1.

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Cut & Run mapping the alteration of H4K20me1 and transcription activity in cultured MuSC upon Suv420h1 depletion.

Project description:H4K20me1/2/3 is dynamically regulated during cell cycle to maintain the genomic stability. Here, we found that depletion of H4K20 di-methyltransferase Suv420h1 in mouse adult MuSCs leads to strong accumulation of H4K20me1 specifically in S phase. In response to it, the transcription activity is increased upon Suv420h1 depletion. To investigate which genomic regions and how the transcription are regulated by Suv420h1 in MuSC, we performed Cut&Run of H4K20me1 and RNA polymerase II Ser2P in control and Suv420h1 inactivated MuSCs.

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DRIP-seq mapping the R-loop formation in cultured Ctrl and Suv420h1 mutant MuSC

Project description:Inactivation of Suv420h1 in MuSC lead to increased transcription and replication collision thereby caused accumulation of RNA:DNA hybrids. This study profiles the R-loops in Ctrl and Suv420h1m mutant MuSCs.

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Project description:Expression data from Suv420h1 knockout and mutant PREX2 expressing MEFs (mouse embryonic fibroblasts)

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