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Physical interaction of estrogen receptor with MnSOD: implication in mitochondrial O2.− upregulation and mTORC2 potentiation in estrogen-responsive breast cancer cells

Oncogene volume 36, pages 1829–1839 (2017)Cite this article

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Abstract

Augmented reactive oxygen species levels consequential to functional alteration of key mitochondrial attributes contribute to carcinogenesis, either directly via oxidative DNA damage infliction or indirectly via activation of oncogenic signaling cascades. We previously reported activation of a key oncogenic signaling cascade via mammalian target of rapamycin (mTOR) signaling complex-2 (mTORC2) owing to estrogen receptor (ER-α)-dependent augmentation of O2.− within the mitochondria of 17-β-estradiol (E2)-stimulated breast cancer cells. Manganese superoxide dismutase (MnSOD) is the principal mitochondrial attribute governing mitochondrial O2.− homeostasis, raising the possibility that its functional alteration could be instrumental in augmenting mitochondrial O2.− levels in breast cancer cells. Here we show ER-dependent transient inhibition of MnSOD catalytic function in breast cancer cells. Catalytic function of MnSOD is tightly regulated at the post-translational level. Post-translational modifications such as phosphorylation, nitration and acetylation represent key regulatory means governing the catalytic function of MnSOD. Acetylation at lysine-68 (K68) inhibits MnSOD catalytic activity and thus represents an important post-translational regulatory mechanism in human cells. Using reciprocal immunoprecipitation and proximity ligation assay, we demonstrate the occurrence of direct physical interaction between ER-α and MnSOD in human breast cancer cells, which in turn was associated with potentiated acetylation of MnSOD at K68. In addition, we also observed diminished interaction of MnSOD with sirtuin-3, the key mitochondrial deacetylase that deacetylates MnSOD at critical K68 and thereby activates it for scavenging O2.−. Consequently, compromised deacetylation of MnSOD at K68 leading to its inhibition and a resultant buildup of O2.− within the mitochondria culminated in the activation of mTORC2. In agreement with this, human breast cancer tissue specimen exhibited a positive correlation between acetyl-MnSODK68 levels and phospho-Ser2481 mTOR levels. In addition to exposing the crosstalk of ER-α with MnSOD post-translational regulatory mechanisms, these data also unravel a regulatory role of ER/MnSOD interaction as an important control switch for redox regulation of ER-α-responsive oncogenic signaling cascades. Furthermore, our study provides a mechanistic link for ER-α-dependent O2.− potentiation and resultant mTORC2 activation in breast cancer cells.

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Acknowledgements

We thank the Director of CDRI for providing the facility and infrastructure. We also thank Dr Kalyan Mitra and Dr Kavita Singh (SAIF) for providing microscopy facilities and Dr AL Vishwakarma (SAIF) for providing flow cytometry facilities. The financial grants received for this study were SR/FT/LS-169/2012 from DST and No. BSC0103 from CSIR India. Mehraj u din alone received fellowship from the Indian Council of Medical Research, New Delhi under ICMR-JRF scheme vide award letter no. 3/1/3/JRF-2011/HRD-69(12035). The institutional manuscript no. for this article is 9324.

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Author notes

  1. R Shrivastava, C Tripathi & S Bhadauria

    Present address: 8Current address: Academy of Scientific and Innovative Research (AcSIR), New Delhi, India.,

  2. M-U-D Lone, K S Baghel and R K Kanchan: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Toxicology, CSIR-Cental Drug Research Institute, Lucknow, India

    M-U-D Lone, K S Baghel, R K Kanchan, R Shrivastava, S A Malik, C Tripathi & S Bhadauria

  2. Department of Surgical Oncology, KGMU, Lucknow, India

    B N Tewari

  3. Department of Clinical and Experimental Medicine, CSIR-CDRI, Lucknow, India

    M P S Negi

  4. Department of Oncosurgery, Lucknow Cancer Institute, Lucknow, India

    V K Garg

  5. Department of Pathology, Lucknow Cancer Institute, Lucknow, India

    M Sharma

  6. Department of Radiotherapy, KGMU, Lucknow, India

    M L B Bhatt

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Correspondence to S Bhadauria.

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Lone, MUD., Baghel, K., Kanchan, R. et al. Physical interaction of estrogen receptor with MnSOD: implication in mitochondrial O2.− upregulation and mTORC2 potentiation in estrogen-responsive breast cancer cells. Oncogene 36, 1829–1839 (2017). https://doi.org/10.1038/onc.2016.346

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