7 results on '"Zdenek Andrysik"'
Search Results
2. Transcriptional Responses to IFN-γ Require Mediator Kinase-Dependent Pause Release and Mechanistically Distinct CDK8 and CDK19 Functions
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Terezia Vcelkova, Heather Bender, Iris Steinparzer, Giulio Superti-Furga, Matthew D. Galbraith, Vitaly Sedlyarov, Lucy Sneezum, Renata Kleinova, Dylan J. Taatjes, Robin D. Dowell, Kevin Eislmayr, Zdenek Andrysik, Jonathan D. Rubin, Joaquín M. Espinosa, Fabian Amman, Florian Wascher, Cecilia B. Levandowski, and Pavel Kovarik
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0303 health sciences ,biology ,Kinase ,RNA polymerase II ,Cell Biology ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Mediator ,Transcription (biology) ,biology.protein ,Cyclin-dependent kinase 8 ,STAT1 ,Kinase activity ,Molecular Biology ,Transcription factor ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Summary Transcriptional responses to external stimuli remain poorly understood. Using global nuclear run-on followed by sequencing (GRO-seq) and precision nuclear run-on sequencing (PRO-seq), we show that CDK8 kinase activity promotes RNA polymerase II pause release in response to interferon-γ (IFN-γ), a universal cytokine involved in immunity and tumor surveillance. The Mediator kinase module contains CDK8 or CDK19, which are presumed to be functionally redundant. We implemented cortistatin A, chemical genetics, transcriptomics, and other methods to decouple their function while assessing enzymatic versus structural roles. Unexpectedly, CDK8 and CDK19 regulated different gene sets via distinct mechanisms. CDK8-dependent regulation required its kinase activity, whereas CDK19 governed IFN-γ responses through its scaffolding function (i.e., it was kinase independent). Accordingly, CDK8, not CDK19, phosphorylates the STAT1 transcription factor (TF) during IFN-γ stimulation, and CDK8 kinase inhibition blocked activation of JAK-STAT pathway TFs. Cytokines such as IFN-γ rapidly mobilize TFs to “reprogram” cellular transcription; our results implicate CDK8 and CDK19 as essential for this transcriptional reprogramming.
- Published
- 2019
3. PO-077 Identification of DHX30 as an inhibitor of the translation of pro-apoptotic mRNAS after P53 activation by nutlin
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Joaquín M. Espinosa, Erik Dassi, Sara Zaccara, Bartolomeo Bosco, Zdenek Andrysik, Annalisa Rossi, Matthew D. Galbraith, Alessandro Quattrone, Alberto Inga, and Dario Rizzotto
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Cloning ,Cancer Research ,chemistry.chemical_compound ,Oncology ,chemistry ,Consensus sequence ,RNA ,Gene silencing ,Luciferase ,Nutlin ,Gene ,Transcription factor ,Cell biology - Abstract
Introduction The transcription factor p53 can be efficiently activated by the small molecule Nutlin-3 without inducing genotoxic stress. Treatment of different cell lines with this small molecule can result in different phenotypes, ranging from cell cycle arrest to apoptosis. HCT116 (colon cancer-derived cells) and SJSA1 (osteosarcoma-derived cells) were used to model the opposite behaviour respectively, by analysing the transcriptional and translational responses after Nutlin-3 treatment. Material and methods Total and polysomal-bound mRNAs were collected and sequenced after 12 hour of 10 uM Nutlin-3 treatment. A bioinformatics analysis of the polysome-enriched mRNAs using Weeder allowed the identification of a 3’UTR motif (‘CG-rich’) which is enriched in the translationally upregulated genes of SJSA1. The effect of the motif on translation was evaluated after cloning its consensus sequence in the 3’UTR of the b-globin gene, which was put downstream the luciferase reporter. The activity of the construct was evaluated after 12 or 24 hours of Nutlin-3. The same consensus was used for a pull-down experiment followed by mass spectrometry to identify proteins interacting with it. Results and discussions RNA-seq data indicate that HCT116 and SJSA1, although sharing almost completely the transcriptional program lead by p53, show almost no overlap at a translation level. SJSA1 present different pro-apoptotic translationally-upregulated genes after Nutlin-3, which have one or more instances of a CG-rich motif in the 3’UTR. The impact of the motif is to enhance the activity of the luciferase reported when cloned in two copies flanking the 3’UTR of the b-globin gene, but only in SJSA1. A pull-down experiment using an RNA bait with the consensus motif was used to identify interactors, among which DHX30 was deeply studied. DHX30 silencing in HCT116 causes: 1) enhanced the activity of the reporter construct after Nutlin; 2) polysomal association of selected mRNAs containing the motif; 3) induction of apoptosis as assessed by Annexin-V staining. In addition, silencing of DHX30 in U2OS cells decreased their survival after Nutlin-3 treatment. Conclusion We show how a p53-dependent transcriptional program can be shaped at a translational level thanks to the action of a CG-rich motif which is enriched in the 3’UTR of some pro-apoptotic mRNAs and that can be bound by DHX30. This protein acts as a translational repressor of mRNAs containing the motif. The exact mechanism and the generalisation of the model are currently being investigated.
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- 2018
4. Autophagy Inhibition Mediates Apoptosis Sensitization in Cancer Therapy by Relieving FOXO3a Turnover
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Jim O'Prey, Zdenek Andrysik, Michael J. Morgan, Brent E. Fitzwalter, Michael Ludwig, Kelly D. Sullivan, Christina G. Towers, Maria Hoh, Joaquín M. Espinosa, Andrew Thorburn, and Kevin M. Ryan
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0301 basic medicine ,Programmed cell death ,Apoptosis ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Puma ,Autophagy ,Humans ,Molecular Biology ,Transcription factor ,biology ,Forkhead Box Protein O3 ,Forkhead Transcription Factors ,Cell Biology ,biology.organism_classification ,030104 developmental biology ,Gene Expression Regulation ,chemistry ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,biology.protein ,Mdm2 ,Growth inhibition ,Apoptosis Regulatory Proteins ,Developmental Biology - Abstract
Macroautophagy (autophagy) is intimately linked with cell death and allows cells to evade apoptosis. This has prompted clinical trials to combine autophagy inhibitors with other drugs with the aim of increasing the likelihood of cancer cells dying. However, the molecular basis for such effects is unknown. Here, we describe a transcriptional mechanism that connects autophagy to apoptosis. The autophagy-regulating transcription factor, FOXO3a, is itself turned over by basal autophagy creating a potential feedback loop. Increased FOXO3a upon autophagy inhibition stimulates transcription of the pro-apoptotic BBC3/PUMA gene to cause apoptosis sensitization. This mechanism explains how autophagy inhibition can sensitize tumor cells to chemotherapy drugs and allows an autophagy inhibitor to change the action of an MDM2-targeted drug from growth inhibition to apoptosis, reducing tumor burden in vivo. Thus, a link between two processes mediated via a single transcription factor binding site in the genome can be leveraged to improve anti-cancer therapies.
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- 2018
5. The aryl hydrocarbon receptor-dependent deregulation of cell cycle control induced by polycyclic aromatic hydrocarbons in rat liver epithelial cells
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Lenka Svihálková-Sindlerová, Cornelia Dietrich, Carsten Weiss, Dagmar Faust, Anne Kranz, Jan Vondráček, Zdenek Andrysik, Alois Kozubík, Miroslav Machala, and Pavel Krcmar
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Health, Toxicology and Mutagenesis ,Cyclin A ,Gene Expression ,Apoptosis ,Cell Cycle Proteins ,Cell Line ,Benz(a)Anthracenes ,Benzo(a)pyrene ,Cytochrome P-450 CYP1A1 ,polycyclic compounds ,Genetics ,Animals ,Rat liver ‘stem-like’ cells ,RNA, Messenger ,Polycyclic Aromatic Hydrocarbons ,RNA, Small Interfering ,Molecular Biology ,Aryl hydrocarbon receptor ,Cell proliferation ,Carcinogen ,Cell Proliferation ,Fluorenes ,Base Sequence ,biology ,Chemistry ,Cell growth ,Cell Cycle ,Cyclin-Dependent Kinase 2 ,Contact inhibition ,Epithelial Cells ,Transfection ,Molecular biology ,Polycyclic aromatic hydrocarbons ,Polycyclic Hydrocarbons, Aromatic ,Rats ,Receptors, Aryl Hydrocarbon ,Biochemistry ,Multiprotein Complexes ,Mutation ,Hepatocytes ,biology.protein ,CDK inhibitor ,Mutagens - Abstract
Disruption of cell proliferation control by polycyclic aromatic hydrocarbons (PAHs) may contribute to their carcinogenicity. We investigated role of the aryl hydrocarbon receptor (AhR) in disruption of contact inhibition in rat liver epithelial WB-F344 'stem-like' cells, induced by the weakly mutagenic benz[a]anthracene (BaA), benzo[b]fluoranthene (BbF) and by the strongly mutagenic benzo[a]pyrene (BaP). There were significant differences between the effects of BaA and BbF, and those of the strongly genotoxic BaP. Both BaA and BbF increased percentage of cells entering S-phase and cell numbers, associated with an increased expression of Cyclin A and Cyclin A/cdk2 complex activity. Their effects were significantly reduced in cells expressing a dominant-negative AhR mutant (dnAhR). Roscovitine, a chemical inhibitor of cdk2, abolished the induction of cell proliferation by BbF. However, neither BaA nor BbF modulated expression of the principal cdk inhibitor involved in maintenance of contact inhibition, p27(Kip1), or pRb phosphorylation. The strongly mutagenic BaP induced apoptosis, a decrease in total cell numbers and significantly higher percentage of cells entering S-phase than either BaA or BbF. Given that BaP induced high levels of Cyclin A/cdk2 activity, downregulation of p27(Kip1) and hyperphosphorylation of pRb, the accumulation of cells in S-phase was probably due to cell proliferation, although S-phase arrest due to blocked replication forks can not be excluded. Both types of effects of BaP were significantly attenuated in dnAhR cells. Transfection of WB-F344 cells with siRNA targeted against AhR decreased induction of Cyclin A induced by BbF or BaP, further supporting the role of AhR in proliferative effects of PAHs. This suggest that activation of AhR plays a significant role both in disruption of contact inhibition by weakly mutagenic PAHs and in genotoxic effects of BaP possibly leading to enhanced cell proliferation. Thus, PAHs may increase proliferative rate and the likelihood of fixation of mutations. This work was partly supported by ECNIS, European Union 6th Framework Program, Priority 5: “Food Quality and Safety” (Contract No. 513943).
- Published
- 2007
6. 447 Acute exposure to polycyclic aromatic hydrocarbons affects multiple cell signaling components in rat liver epithelial cell line
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Miroslav Machala, Brad L. Upham, Jiří Neča, Zdenek Andrysik, Katerina Pencikova, Luděk Bláha, Jan Vondráček, and Petra Kapplová
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Cell signaling ,medicine.anatomical_structure ,Biochemistry ,Chemistry ,Rat liver ,Acute exposure ,medicine ,General Medicine ,Line (text file) ,Toxicology ,Epithelium ,Cell biology - Published
- 2003
7. 449 AHR-activating polycyclic aromatic hydrocarbons induce a release from contact inhibition or apoptosis in rat liver epithelial cell line
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Katerina Chramostova, Zdenek Andrysik, Miroslav Machala, Alois Kozubík, Jan Vondráček, Karel Souček, and B. Vojtesek
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medicine.anatomical_structure ,Apoptosis ,Chemistry ,Rat liver ,medicine ,Contact inhibition ,Organic chemistry ,General Medicine ,Line (text file) ,Toxicology ,Epithelium ,Cell biology - Published
- 2003
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