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DiazMeco_Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer_set1

ABSTRACT: Identification of interacting partners from immuno-precipitated HA-EZH2 samples. Endogenous Co-IP was performed using 1.5 mg of nuclear lysate with anti-HA antibody after transfecting HEK 293T cells with HA-EZH2 plasmid. Two IP-ed samples were submitted to the core for analysis, HEK293T-sgC and sgPRKCI.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Maria T. Diaz-Meco Conde

PROVIDER: MSV000094704 | MassIVE | Tue May 07 12:06:00 BST 2024

SECONDARY ACCESSION(S): PXD052091

REPOSITORIES: MassIVE

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DiazMeco_Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer_set3

Project description:Identification of phosphopeptides EZH2 in control and kinase-deficient cells. The HA-EZH2 plasmid was transfected in HEK 293T cells, and HA-EZH2 was subsequently purified using HA peptide. The elution was performed with 1mg/mL HA-peptide in Tris buffered saline (TBS with 150mM NaCl), pH 7.5 in a final volume of 100 microL and a concentration of 150-250 ng/microL. Two samples were sent to the facility for analysis: HEK293T-sgC and sgPRKCI.

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Project description:Identification of phosphopeptides in EZH2 upon PRKCI treatment (kinase). In vitro kinase reactions were performed in buffer containing 25 mM HEPES, 15 mM MgCl2, and 1mM DTT. Two samples were sent to the facility for analysis, EZH2, and EZH2+PRKCI.

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Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer (CUT&RUN)

Project description:Resistance to therapy is a major challenge in castration-resistant prostate cancer (CRPC). Lineage plasticity towards a neuroendocrine phenotype enables CRPC to adapt and survive targeted-therapies. However, the molecular mechanisms of epigenetic reprogramming during this process are still poorly understood. Here we show that the protein kinase PKCl/i phosphorylates the epigenetic regulator enhancer of zeste homologue 2 (EZH2) to regulate its proteasomal degradation and maintain EZH2 as part of the canonical polycomb repressive complex (PRC2). Loss of PKCl/i promotes a switch during enzalutamide treatment to a non-canonical EZH2 cistrome that triggers the transcriptional activation of the translational machinery to induce a transforming growth factor b (TGFb) resistance program. The increased reliance on protein synthesis creates a synthetic vulnerability in PKCl/i-deficient CRPC.

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Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer (scRNA-Seq)

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Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer (CUT&RUN) II

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Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer (RNA-Seq polysome)

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Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer

Project description:This SuperSeries is composed of the SubSeries listed below.

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PAR-CLIP for HA-tagged EZH2 in mouse embryonic stem cells

Project description:This experiment was designed to indentify RNAs making direct contact with EZH2 in mouse embryonic stem cells E14 with an integrated transgene encoding HA-EZH2 were pulsed with 4-SU, irradiated with UV, and subjected to HA immunoprecipitation.

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