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Importance of Transcriptional coactivator PC4 in p53-dependent Genotoxic Stress Response

ABSTRACT: In order to get an idea of involvement of PC4 in regulation of p53-dependent gene expression, we carried out global gene expression profiling upon treatment with R6-NLS-p53(380–386-3Ac). R6-NLS-p53(380–386-3Ac) is a small peptide of 20 residues derived from p53 C-terminal domain. This peptide can successfully block the cellular p53-PC4 interactions. Lung cancer line A549 which contain both wild type p53 and PC4 were used for this study. The cells were treated with 25µM R6-NLS-p53(380–386-3Ac) for two different time points (12 h and 24 h) and the gene expression profile was analyzed by microarray analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE116020 | GEO | 2019/06/21

REPOSITORIES: GEO

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Project description:Spermatogonial stem cells (SSCs) can spontaneously dedifferentiate into embryonic stem cell (ESC)-like cells, which are designated as multipotent SSCs (mSSCs), without ectopic expression of reprogramming factors. SSCs express key OSKM reprogramming factors at some levels, and do not require ectopic expression of any gene for the acquisition of pluripotency during reprogramming to mSSCs. Therefore, we reasoned that additional factors are required to regulate SSC reprogramming. In this study, we first compared the expression of reprogramming signature genes among somatic cells, iPSC, SSCs, mSSCs, and partially reprogramed cells, and found that they appear to have similar pluripotency states, whereas their transcriptional program differs. We developed a systems biology approach to prioritise genes for pluripotency regulatory factors by integrating transcriptome and interactome data on the genome-wide functional network. Then, we performed a series of systematic gene prioritisation steps and identified 53 candidates, which included some known reprogramming factors. We experimentally validated one particular candidate, Positive cofactor 4 (Pc4), which was expressed in PSCs and yielded a positive RNA interference (RNAi) response in an Oct4 reporter assay. We demonstrated that Pc4 enhanced the efficiency of OSKM-mediated reprogramming by promoting the transcriptional activity of key pluripotency factors, and by regulating the expression of many protein- and miRNA-encoding genes involved in reprogramming and somatic cell-specific genes. Pc4-overexpressing mESC lines were established by Venus (YFP)-expressing lentiviral transfection. The mESCs were split at a density of 2 Â´ 104 cells onto fresh MEF feeder cells seeded into a 6 well dish (containing mESC growth medium) with virus particles, and 25 Î¼g/ml polybrene (Sigma Aldrich) was added. After 24 h, the medium was replaced with fresh growth medium. After 4 days later, mESC colonies expressing YFP were picked and replated. Three different Pc4-overexpressing mESC lines were established.

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Importance of Transcriptional coactivator PC4 in p53-dependent Genotoxic Stress Response

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