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Transcription factor Ets-2 regulates the expression of key lymphotropic factors

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Abstract

Transcription factor Ets-2 downregulates the expression of cytokine genes and HIV-1 in resting T-cells. Herein, we studied whether Ets-2 regulates the expression of lymphotropic factors (LFs) NFAT2, NF-κΒ/p65, c-Jun, c-Fos, which regulate the activation/differentiation of T-cells, and kinase CDK10, which controls Ets-2 degradation and repression activity. In silico analysis revealed Ets-2 binding sites on the promoters of NFAT2, c-Jun, c-Fos. The T-cell lines Jurkat (models T-cell signaling/activation) and H938 (contains the HIV-1-LTR) were transfected with an Ets-2 overexpressing vector, in the presence/absence of mitogens. mRNA and protein levels were assessed by qPCR and Western immunoblotting, respectively. Ets-2 overexpression in unstimulated Jurkat increased NFAT2 and c-Jun mRNA/protein, c-Fos mRNA and NF-κΒ/p65 protein, and decreased CDK10 protein. In unstimulated H938, Ets-2 upregulated NFAT2, c-Jun and CDK10 mRNA/protein and NF-κΒ/p65 protein. In stimulated Jurkat, Ets-2 increased NFAT2, c-Jun and c-Fos mRNA/protein and decreased CDK10 mRNA/protein. In stimulated H938 Ets-2 increased NFAT2, c-Jun and c-Fos protein and reduced CDK10 protein levels. Furthermore, Ets-2 overexpression modulated the expression of pro- and anti-apoptotic genes in both cell lines. Ets-2 upregulates the expression of key LFs involved in the activation of cytokine genes or HIV-1 in T-cells, either through its physical interaction with gene promoters or through its involvement in signaling pathways that directly impact their expression. The effect of Ets-2 on CDK10 expression in H938 vs Jurkat cells dictates that, additionally to Ets-2 degradation, CDK10 may facilitate Ets-2 repression activity in cells carrying the HIV-1-LTR, contributing thus to the regulation of HIV latency in virus-infected T-cells.

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Funding

This research was co-financed by Greece and the European Union (European Social Fund- ESF) through the Operational Program “Human Resources Development, Education and Lifelong Learning” in the context of the project “Strengthening Human Resources Research Potential via Doctorate Research” (MIS-5000432), implemented by the State Scholarships Foundation (ΙΚΥ) (to PD), the Operational Program «Human Resources Development, Education and Lifelong Learning» in the context of the project "Reinforcement of Postdoctoral Researchers—2nd Cycle" (MIS-5033021), implemented by the State Scholarships Foundation (ΙΚΥ) (to IA), and GILEAD-ASKLEPIOS grants for 2018 and 2019 (to AM).

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Authors and Affiliations

  1. Laboratory of Immunohematology, Division of Hematology, Department of Internal Medicine, Medical School, University of Patras, 26500, Patras, Greece

    Panagiota Davoulou, Ioanna Aggeletopoulou, Ioannis Panagoulias, Tassos Georgakopoulos & Athanasia Mouzaki

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  1. Panagiota Davoulou
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  2. Ioanna Aggeletopoulou
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  5. Athanasia Mouzaki
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AM conceived and coordinated the study; PD, IA, IP performed the experiments, analyzed the data and prepared the figures; TG and IP advised on methodology; PD, IA, TG and AM wrote the manuscript.

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Correspondence to Athanasia Mouzaki.

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Davoulou, P., Aggeletopoulou, I., Panagoulias, I. et al. Transcription factor Ets-2 regulates the expression of key lymphotropic factors. Mol Biol Rep 47, 7871–7881 (2020). https://doi.org/10.1007/s11033-020-05865-x

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