Abstract
Knockout of heat shock protein Hsp72 was shown to promote chromosomal instability and increase radiation sensitivity of mouse fibroblasts. Here, we report that downregulation of Hsp72 in human tumor cells leads to suppression of a specific branch of the DNA damage response (DDR) that facilitates DNA repair following genotoxic insults, that is, reduced accumulation of the phosphorylated form of histone H2AX (γH2AX). This inhibition was due to decreased expression of H2AX as well as higher rate of γH2AX dephosphorylation. Formation of γH2AX and MDC1 radiation-induced foci was impaired in Hsp72-depleted cells, which in turn enhanced DNA damage, resulting in sensitization of cells to γ-radiation and doxorubicin. These effects of Hsp72 knockdown were dependent on activation of the p53/p21-signaling pathway. Overall, permanent activation of the p53/p21 signaling in Hsp72-depleted cells specifically impaired the γH2AX pathway of the DDR, enhanced DNA damage following genotoxic insults, and led to further stimulation of the p53/p21 pathway, thus creating a positive feedback loop. The resulting strong induction of p21 precipitated senescence following exposure to DNA-damaging agents, thus accounting for higher sensitivity of cells to genotoxic stresses.
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References
Bakkenist CJ, Kastan MB . (2004). Initiating cellular stress. Responses Cell 118: 9–17.
Bonner WM, Redon CE, Dickey JS, Nakamura AJ, Sedelnikova OA, Solier S et al. (2008). GammaH2AX and cancer. Nat Rev Cancer 2008 8: 957–967.
Cao C, Shinohara ET, Subhawong TK, Geng L, Woon Kim K, Albert JM et al. (2006). Radiosensitization of lung cancer by nutlin, an inhibitor of murine double minute 2. Mol Cancer Ther 5: 411–417.
Chowdhury D, Keogh M-C, Ishii H, Peterson C, Buratowski S, Lieberman J . (2005). g-H2AX Dephosphorylation by protein phosphatase 2A facilitates DNA double-strand break repair. Molecular Cell 20: 801–809.
Ciocca DR, Calderwood SK . (2005). Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. Cell Stress Chaperones 10: 86–103.
Efeyan A, Ortega-Molina A, Velasco-Miguel S, Herranz D, Vassilev LT, Serrano M . (2007). Induction of p53-dependent senescence by the MDM2 antagonist nutlin-3a in mouse cells of fibroblast origin. Cancer Res 67: 7350–7357.
Gabai VL, O'Callaghan-Sunol C, Meng L, Sherman MY, Yaglom J . (2008). Triggering senescence programs suppresses chk1 kinase and sensitizes cells to genotoxic stresses. Cancer Res 68: 1834–1842.
Gorbunova V, Seluanov A, Mao Z, Hine C . (2007). Changes in DNA repair during aging. Nucl Acids Res 35: 7466–7474.
Hunt CR, Dix DJ, Sharma GG, Pandita RK, Gupta A, Funk M et al. (2004). Genomic instability and enhanced radiosensitivity in Hsp70.1- and Hsp70.3-deficient mice. Mol Cell Biol 24: 899–911.
Kabakov AE, Malyutina YV, Latchman DS . (2006). Hsf1-mediated stress response can transiently enhance cellular radioresistance. Radiat Res 165: 410–423.
Li L, Zou L . (2005). Sensing, signaling, and responding to DNA damage: organization of the checkpoint pathways in mammalian cells. J Cell Biochem 94: 298–306.
Lou Z, Minter-Dykhouse K, Franco S, Gostissa M, Rivera MA, Celeste A et al. (2006). MDC1 maintains genomic stability by participating in the amplification of ATM-dependent DNA damage signals. Mol Cell 21: 187–200.
Niida H, Nakanishi M . (2006). DNA damage checkpoints in mammals. Mutagenesis 21: 3–9.
O′Callaghan-Sunol C, Gabai VL, Sherman MY . (2007). Hsp27 modulates p53 signaling and suppresses cellular senescence. Cancer Res 67: 11779–11788.
O′Callaghan-Sunol C, Gabai VL . (2007). Heat Shock Proteins in Cancer. Calderwood S, Sherman My, Ciocca DR (eds). Springer: Dordrecht, Netherlands, pp 169–190.
Park J, Park E, Lee H, Kim S, Hur S, Imbalzano A et al. (2006). Mammalian SWI/SNF complexes facilitate DNA double-strand break repair by promoting gamma-H2AX induction. EMBO J 25: 3986–3997.
Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn S . (2004). Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Ann Rev Biochem 73: 39–85.
Sedelnikova OA, Horikawa I, Zimonjic DB, Popescu NC, Bonner WM, Barrett JC . (2004). Senescing human cells and ageing mice accumulate DNA lesions with unrepairable double-strand breaks. Nat Cell Biol 6: 168–170.
Sedelnikova OA, Horikawa I, Redon C, Nakamura A, Zimonjic DB, Popescu NC . (2008). Delayed kinetics of DNA double-strand break processing in normal and pathological aging. Aging Cell 7: 89–100.
Stommel JW, Wahl GM . (2004). Accelerated MDM2 auto-degradation induced by DNA-damage kinases is required for p53 activation. EMBO J 23: 1547–1556.
Vassilev LT . (2007). MDM2 inhibitors for cancer therapy. Trends Mol Med 13: 23–31.
Wang R-H, Sengupta K, Li C, Kim H-S, Cao L, Xiao C et al. (2008). Impaired DNA damage response, genome instability, and tumorigenesis in sirt1 mutant mice. Cancer Cell 14: 312–323.
Yaglom J, O′Callaghan-Sunol C, Gabai V, Sherman MY . (2003). Inactivation of dual-specificity phosphatases is involved in the regulation of extracellular signal-regulated kinases by heat shock and Hsp72. Mol Cell Biol 23: 3813–3824.
Yaglom JA, Gabai VL, Sherman MY . (2007). High levels of heat shock protein hsp72 in cancer cells suppress default senescence pathways. Cancer Res 67: 2373–2381.
Acknowledgements
This work was supported by American Cancer Society Institutional Grant (VLG); NIH grant (MYS) and Grunnenbaum Award (JAY).
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Gabai, V., Sherman, M. & Yaglom, J. HSP72 depletion suppresses γH2AX activation by genotoxic stresses via p53/p21 signaling. Oncogene 29, 1952–1962 (2010). https://doi.org/10.1038/onc.2009.480
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DOI: https://doi.org/10.1038/onc.2009.480
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