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- Title
Transcriptome Analysis of Cells Exposed to Actinomycin D and Nutlin-3a Reveals New Candidate p53-Target Genes and Indicates That CHIR-98014 Is an Important Inhibitor of p53 Activity.
- Authors
Łasut-Szyszka, Barbara; Małachowska, Beata; Gdowicz-Kłosok, Agnieszka; Krześniak, Małgorzata; Głowala-Kosińska, Magdalena; Zajkowicz, Artur; Rusin, Marek
- Abstract
Co-treatment with actinomycin D and nutlin-3a (A + N) strongly activates p53. Previously we reported that CHIR-98014 (GSK-3 kinase inhibitor), acting in cells exposed to A + N, prevents activation of TREM2-an innate immunity and p53-regulated gene associated with Alzheimer's disease. In order to find novel candidate p53-target genes and genes regulated by CHIR-98014, we performed RNA-Seq of control A549 cells and the cells exposed to A + N, A + N with CHIR-98014 or to CHIR-98014. We validated the data for selected genes using RT-PCR and/or Western blotting. Using CRISPR/Cas9 technology we generated p53-deficient cells. These tools enabled us to identify dozens of candidate p53-regulated genes. We confirmed that p53 participates in upregulation of BLNK, APOE and IRF1. BLNK assists in activation of immune cells, APOE codes for apolipoprotein associated with Alzheimer's disease and IRF1 is activated by interferon gamma and regulates expression of antiviral genes. CHIR-98014 prevented or inhibited the upregulation of a fraction of genes stimulated by A + N. Downregulation of GSK-3 did not mimic the activity of CHIR-98014. Our data generate the hypothesis, that an unidentified kinase inhibited by CHIR-98014, participates in modification of p53 and enables it to activate a subset of its target genes, e.g., the ones associated with innate immunity.
- Subjects
DACTINOMYCIN; APOLIPOPROTEIN E4; CELL analysis; TRANSCRIPTOMES; P53 antioncogene; INTERFERON gamma; GENES
- Publication
International Journal of Molecular Sciences, 2021, Vol 22, Issue 20, p11072
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms222011072