Unknown,Transcriptomics,Genomics,Proteomics

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VuDREB2A activated transcription in wild type Arabidopsis: VuDREB2A overexpressor lines vs. wild type Arabidopsis

ABSTRACT: Transcriptional profiling of Arabidopsis overexpressing VuDREB2A (full length/truncated) compared to wild type. Unstressed three week-old whole seedlings were used. Goal was to search the genes upregulated byVuDREB2A in Arabidopsis. Two-condition experiment, VuDREB2A (full length/truncated) overexpressing lines v.s. wild type. Biological replicates: 2 transgenic replicates, 2 wild type replicates

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Ayan Sadhukhan

PROVIDER: E-GEOD-53113 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Reactivation of androgen receptor (AR) may drive recurrent prostate cancer in castrate patients. Ack1 tyrosine kinase is overexpressed in prostate cancer and promotes castrate resistant xenograft tumor growth and enhances androgen target gene expression and AR recruitment to enhancers. Ack1 phosphorylates AR at Tyr-267 in the N-terminal transactivation domain. In this study, the role of this phosphorylation site was investigated by characterizing the phosphorylation site mutant in the context of full length and truncated AR lacking the ligand-binding domain. The Y267F mutant showed decreased transactivation of reporters. Expression of wild type full length and truncated AR in LNCaP cells increased cell proliferation in androgen-depleted conditions and increased colony formation. However, the Y267F mutant of full length and truncated AR was defective in stimulating cell proliferation. The full length AR Y267F mutant was defective in nuclear translocation induced by androgen or Ack1 kinase. The truncated AR was constitutively localized to the nucleus. Chromatin immunoprecipitation analysis showed that it was recruited to the target enhancers without androgen. The truncated Y267F AR mutant did not exhibit constitutive nuclear localization and androgen enhancer binding activity. Expression of AR responsive genes in cells expressing truncated AR wt and AR-Y267F mutant under androgen deprived conditions was assessed by microarray gene expression analysis. The AR pathway score was increased in cells expressing truncated AR wt and decreased in cells expressing truncated AR-Y267F. These results support the concept that phosphorylation of Tyr-267 is required for AR nuclear translocation and recruitment and DNA binding and transcription of AR responsive genes. Gene expression profiling was performed using RNA samples from LNCaP cells expressing truncated AR-wt and truncated AR-Y267F growing in androgen deprived conditions. The RNA sample from vector control cells were used as reference sample. Two biological replicates were used per each cell line.

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VuDREB2A activated transcription in wild type Arabidopsis: VuDREB2A overexpressor lines vs. wild type Arabidopsis

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