c-Myc Suppression of DNA Double-strand Break Repair<sup><a href="#FN1" rid="FN1" class=" fn">1</a>,</sup><sup><a href="#FN2" rid="FN2" class=" fn">2</a></sup>

Zhaozhong Li

Taofeek Owonikoko

Shi Sun

Suresh Ramalingam

Supplementary Figures and Tables:

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^{Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA}

^{Department of Hematology and Medical Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA}

^{Department of Biochemistry, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA}

^{Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha, Hunan, China}

Address all correspondence to: Xingming Deng, MD, PhD, Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA 30322. E-mail: ude.yrome@4gnedx

Received 2012 Aug 1; Revised 2012 Oct 13; Accepted 2012 Oct 16.

Abstract

c-Myc is a transcriptional factor that functions as a central regulator of cell growth, proliferation, and apoptosis. Overexpression of c-Myc also enhances DNA double-strand breaks (DSBs), genetic instability, and tumorigenesis. However, the mechanism(s) involved remains elusive. Here, we discovered that γ-ray ionizing radiation-induced DSBs promote c-Myc to form foci and to co-localize with γ-H2AX. Conditional expression of c-Myc in HO15.19 c-Myc null cells using the Tet-Off/Tet-On inducible system results in down-regulation of Ku DNA binding and suppressed activities of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and DNA end-joining, leading to inhibition of DSB repair and enhanced chromosomal and chromatid breaks. Expression of c-Myc reduces both signal and coding joins with decreased fidelity during V(D)J recombination. Mechanistically, c-Myc directly interacts with Ku70 protein through its Myc box II (MBII) domain. Removal of the MBII domain from c-Myc abrogates its inhibitory effects on Ku DNA binding, DNA-PKcs, and DNA end-joining activities, which results in loss of c-Myc's ability to block DSB repair and V(D)J recombination. Interestingly, c-Myc directly disrupts the Ku/DNA-PKcs complex in vitro and in vivo. Thus, c-Myc suppression of DSB repair and V(D)J recombination may occur through inhibition of the nonhomologous end-joining pathway, which provides insight into the mechanism of c-Myc in the development of tumors through promotion of genomic instability.

Abstract

Acknowledgments

We are grateful to Frederick W. Alt (Harvard Medical School) for kindly providing the pJH200, pJH290, RAG1, and RAG2 constructs, Shigemi Matsuyama (Case Western Reserve University) for Flag-tagged Ku70 mutants, and John M. Sedivy (Brown University) for HO15.19 c-Myc null cells. We also thank Anthea Hammond for editing the manuscript.

Acknowledgments

Abbreviations

HR	homologous recombination
NHEJ	nonhomologous end-joining
DSB	DNA double-strand break
PFGE	pulsed-field gel electrophoresis
siRNA	small interfering RNA
RNAi	RNA interference
DOX	doxycycline
DNA-PKcs	DNA-dependent protein kinase catalytic subunit

Abbreviations

Footnotes

^{This work was supported by National Cancer Institute, National Institutes of Health grants R01CA112183 and R01CA136534, by Flight Attendant Medical Research Institute Clinical Innovator Awards, and by National Aeronautics and Space Administration grant NNX12AC30G. The authors declare no conflict of interest.}

^{This article refers to supplementary materials, which are designated by}Figures W1 to W5 and are available online at www.neoplasia.com.

Footnotes

Supplemental References

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