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Inhibitor of growth 1 (ING1) acts at early steps of multiple DNA repair pathways

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Abstract

ING proteins are tumor suppressors involved in the regulation of gene transcription, cell cycle arrest, apoptosis, and senescence. Here, we show that ING1b expression is upregulated by several DNA-damaging agents, in a p53-independent manner. ING1b stimulates DNA repair of a variety of DNA lesions requiring activation of multiple DNA repair pathways. Moreover, Ing1 −/− cells showed impaired genomic DNA repair after H2O2 and neocarzinostatin treatment and this defect was reverted by overexpression of ING1b. Two tumor-derived ING1 mutants failed to promote DNA repair highlighting the physiological importance of the integrity of the PHD domain for ING1b DNA repair activity and suggesting a role in the prevention of tumor progression. Ing−/− cells showed higher basal levels of γ-H2AX and, upon DNA damage, γ-H2AX increase was greater and with faster kinetics compared to wild-type cells. Chromatin relaxation by Trichostatin A led to an exacerbated damage signal in both types of cells, but this effect was dependent on Ing1 status, and more pronounced in wild-type cells. Our results suggest that ING1 acts at early stages of the DNA damage response activating a variety of repair mechanisms and that this function of ING1 is targeted in tumors.

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Abbreviations

ING1:

Inhibitor of growth 1

DDR:

DNA damage response

NCS:

Neocarzinostatin

UDS:

Unscheduled DNA synthesis

PHD:

Plant homeodomain

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Acknowledgments

This study was supported by research grants from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Agencia Nacional de Promoción Científica y Tecnológica (ANPCYT), and Universidad de Buenos Aires to EC and Spanish Ministry of Science and Innovation to IP (SAF2009-09031). We thank Dr. Luciana E. Giono for help with the manuscript.

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Author notes

  1. Camino Menéndez

    Present address: Spanish National Cancer Research Centre (CNIO), Melchor Fernández Almagro, 3, 28029, Madrid, Spain

Authors and Affiliations

  1. Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Buenos Aires, Argentina

    Julieta M. Ceruti, María F. Ogara & Eduardo T. Cánepa

  2. Instituto de Investigaciones Biomédicas “Alberto Sols” CSIC-UAM, Arturo Duperier, 4, 28029, Madrid, Spain

    Camino Menéndez & Ignacio Palmero

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  1. Julieta M. Ceruti
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  2. María F. Ogara
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  3. Camino Menéndez
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  4. Ignacio Palmero
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Correspondence to Ignacio Palmero or Eduardo T. Cánepa.

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11010_2013_1601_MOESM1_ESM.tif

ESM 1 ING1b improves DNA repair in response to UV. HeLa cells were transfected with a vector encoding wild-type ING1b (ING wt) or mutants ING1bC215S (ING 215), ING1bK270 N (ING 270) along with pBabePuro. Twenty-four hours after transfection regular medium was substituted for arginine-free medium containing 1 % FBS and cells were incubated for additional 60 h in presence of puromycin. Resistant cells were treated with 8 mJ/cm2 UV and incubated with 10 mCi [3H] thymidine for 10 h. Cell lysates were tested for genomic DNA repair by UDS assay. Bars represent the mean ± S.E. of three experiments performed in triplicate. Student t test was used to compare samples (*P < 0.05). (TIFF 1645 kb)

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Ceruti, J.M., Ogara, M.F., Menéndez, C. et al. Inhibitor of growth 1 (ING1) acts at early steps of multiple DNA repair pathways. Mol Cell Biochem 378, 117–126 (2013). https://doi.org/10.1007/s11010-013-1601-2

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