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Human EHMT2/G9a activates p53 through methylation-independent mechanism

Oncogene volume 36, pages 922–932 (2017)Cite this article

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Abstract

p53 is a critical tumor suppressor in humans. It functions mostly as a transcriptional factor and its activity is regulated by numerous post-translational modifications. Among different covalent modifications found on p53 the most controversial one is lysine methylation. We found that human G9a (hG9a) unlike its mouse orthologue (mG9a) potently stimulated p53 transcriptional activity. Both ectopic and endogenous hG9a augmented p53-dependent transcription of pro-apoptotic genes, including Bax and Puma, resulting in enhanced apoptosis and reduced colony formation. Significantly, shRNA-mediated knockdown of hG9a attenuated p53-dependent activation of Puma. On the molecular level, hG9a interacted with histone acetyltransferase, p300/CBP, resulting in increased histone acetylation at the promoter of Puma. The bioinformatics data substantiated our findings showing that positive correlation between G9a and p53 expression is associated with better survival of lung cancer patients. Collectively, this study demonstrates that depending on the cellular and organismal context, orthologous proteins may exert both overlapping and opposing functions. Furthermore, this finding has important ramifications on the use of G9a inhibitors in combination with genotoxic drugs to treat p53-positive tumors.

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Abbreviations

DAPI:

4,6 diamidino- 2-phenylindole

GLP:

G9a-like protein

HATs:

histone acetyltransferases

HMTs:

histone methyltransferases

hG9a:

human G9a

hp53:

human p53

KMT:

lysine methyltransferase

mG9a:

mouse G9a

mp53:

mouse p53

PTMs:

post-translational modificaitions.

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Acknowledgements

We thank Dr O. Binda (Newcastle University, UK), Dr MR. Stallcup (University of Southern California, USA), Dr C-M Chiang (University of Texas Southwestern Medical Center, USA), Dr SB Seo (Chung-Ang University, Korea), Dr P. Muller (MRC Toxicology Unit, UK) for providing various constructs and reagents. We also thank S Cowley lab (University of Leicester, UK) for providing E14 cells. This work was supported by grants from RSF 14-15-00816 to (E.V. and N.B.); M.R. was supported by a Doctoral Studentship from the Kurdistan Regional Government (Iraq); A.V.A. was supported by MCB RAS programme; G.M. acknowledges the grant support from Medical Research Council, AIRC 2014 IG15653, AIRC 5xmille MCO9979, AIRC 2011 IG11955 and Fondazione Roma malattie Non trasmissibili Cronico-Degenerative (NCD).

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Authors and Affiliations

  1. Department of Biochemistry, University of Leicester, Leicester, UK

    M Rada, L Lezina, D Marouco & S Macip

  2. Institute of Cytology, Saint-Petersburg, Russia

    E Vasileva, L Lezina, A V Antonov & N A Barlev

  3. MRC Toxicology Unit, Leicester, UK

    A V Antonov & G Melino

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  1. M Rada
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Correspondence to N A Barlev.

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Rada, M., Vasileva, E., Lezina, L. et al. Human EHMT2/G9a activates p53 through methylation-independent mechanism. Oncogene 36, 922–932 (2017). https://doi.org/10.1038/onc.2016.258

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