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Orphan receptor NR4A3 is a novel target of p53 that contributes to apoptosis

Oncogene volume 38, pages 2108–2122 (2019)Cite this article

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Abstract

Major tumor suppressor and transcription factor p53 coordinates expression of many genes hence affecting critical cellular functions including cell cycle, senescence, and apoptosis. The NR4A family of orphan receptors (NR4A1-3) belongs to the superfamily of nuclear receptors. They regulate genes involved in proliferation, cell migration, and apoptosis. In this study, we report an identification of NR4A3 as a direct transcriptional target of p53. Using various techniques, we showed that p53 directly bound the promoter of NR4A3 gene and induced its transcription. Functionally, over-expression of NR4A3 attenuated proliferation of cancer cells and promoted apoptosis by augmenting the expression of pro-apoptotic genes, PUMA and Bax. Knockdown of NR4A3 reversed these phenotypes. Importantly, NR4A3 exhibited tumor suppressive functions both in p53-dependent and independent manner. In addition, NR4A3 physically interacted with an anti-apoptotic Bcl-2 protein hence sequestering it from blunting apoptosis. These observations were corroborated by the bioinformatics analysis, which demonstrated a correlation between high levels of NR4A3 expression and better survival of breast and lung cancer patients. Collectively, our studies revealed a novel transcriptional target of p53, NR4A3, which triggers apoptosis and thus likely has a tumor suppressive role in breast and lung cancers.

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Acknowledgements

O.F., A.P. and E.V. carried out chromatin immunoprecipitation, Luciferase assay, quantitative PCR, cell-cycle analysis, colony formation assay, immunofluorescence, wound-healing and real-time cell migration assay and acknowledge the support from RCF grant 18-75-10076. A.D., O.S., N.B. carried out annexin V, co-immunoprecipitation interaction assay and the bioinformatics analysis and acknowledge the support from RFBR grant 18-29-09144. We appreciate Dr Mikhal Maliewitz (MRC Toxicology, Leicester) for a gift of NR4A1 and NR4A2 antibodies.

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Authors and Affiliations

  1. Gene Expression Program, Institute of Cytology, Saint-Petersburg, Russia, 194064

    Olga Fedorova, Alexey Petukhov, Alexandra Daks, Oleg Shuvalov, Tatyana Leonova, Elena Vasileva, Nikolai Aksenov & Nikolai A. Barlev

  2. Almazov National Medical Research Centre, St. Petersburg, Russia, 197341

    Alexey Petukhov

  3. MRC Toxicology Unit, Leicester, LE1 9HN, UK

    Gerry Melino

  4. Moscow Institute of Technology and Physics, Dolgoprudny, Moscow Region, Russia, 141700

    Nikolai A. Barlev

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  1. Olga Fedorova
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Fedorova, O., Petukhov, A., Daks, A. et al. Orphan receptor NR4A3 is a novel target of p53 that contributes to apoptosis. Oncogene 38, 2108–2122 (2019). https://doi.org/10.1038/s41388-018-0566-8

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