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The nuclear cofactor RAC3/AIB1/SRC-3 enhances Nrf2 signaling by interacting with transactivation domains

Oncogene volume 32, pages 514–527 (2013)Cite this article

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Abstract

Nuclear factor erythroid 2-related factor 2 (Nrf2, NM 006164, 605 AA) is essential for the antioxidant responsive element (ARE)-mediated expression of a group of detoxifying antioxidant genes that detoxify carcinogens and protect against oxidative stress. Several proteins have been identified as Nrf2-interacting molecules. In this study, we found that the overexpression of receptor-associated coactivator 3 (RAC3)/AIB-1/steroid receptor coactivator-3, a nuclear coregulator and oncogene frequently amplified in human breast cancers, induced heme oxygenase-1 (HO-1) through Nrf2 transactivation in HeLa cells. Next, we determined the interaction between RAC3 and Nrf2 proteins using a co-immunoprecipitation assay and fluorescence resonance energy transfer analysis. The results showed that RAC3 bound directly to the Nrf2 protein in the nucleus. Subsequently, we identified the interacting domains of Nrf2 and RAC3 using a glutathione S-transferase pull-down assay. The results showed that both the N-terminal RAC3-pasB and C-terminal RAC3-R3B3 domains were tightly bound to the Neh4 and Neh5 transactivation domains. Furthermore, chromatin immunoprecipitation showed that RAC3 bound tightly to the ARE enhancer region of the HO-1 promoter via Nrf2 binding. These data suggest that Nrf2 activation is modulated and directly controlled through interactions with the RAC3 protein in HeLa cells.

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Abbreviations

ACTR:

activator of retinoid and thyroid receptors

AIB-1:

amplified in breast cancer-1 protein

ARE:

antioxidant responsive element

GST:

glutathione S-transferase

HO-1:

heme oxygenase-1

Keap1:

Kelch-like ECH-associated protein 1

NCoA-3:

nuclear receptor coactivator-3

Neh:

Nrf2-ECH homology

Nrf2:

nuclear factor erythroid 2-related factor 2

pCIP:

CBP-interacting protein

RAC3:

receptor-associated coactivator 3

SRC-3:

steroid receptor coactivator protein-3

TAD:

transactivation domain

TRAM-1:

thyroid hormone receptor activator molecule-1

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Acknowledgements

We thank Dr Jianjie Ma (Department of Physiology and Biophysics, University of Medicine and Dentistry of New Jersey, Newark, New Jersey, USA) for help with confocal microscopy. This study was supported by NIH Grant R01 NIH R01-CA-94828.

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  1. J-H Kim

    Present address: 4Current address: Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.,

Authors and Affiliations

  1. Graduate Program in Pharmaceutical Science, Rutgers, The State University of New Jersey, Piscataway, NJ, USA

    J-H Kim & A N Kong

  2. Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ, USA

    J-H Kim, S Yu & A N Kong

  3. Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ, USA

    J D Chen

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Correspondence to A N Kong.

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Kim, JH., Yu, S., Chen, J. et al. The nuclear cofactor RAC3/AIB1/SRC-3 enhances Nrf2 signaling by interacting with transactivation domains. Oncogene 32, 514–527 (2013). https://doi.org/10.1038/onc.2012.59

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