Abstract
Cellular senescence is important for the maintenance of tissue homeostasis, and has recently been shown to pose a natural barrier to tumorigenesis. The E3 ubiquitin ligase, E6AP, has been linked to a number of protein regulators of the cell cycle as well as the cellular stress response. We therefore explored the role of E6AP in the cellular response to stress. We found that mouse embryo fibroblasts (MEFs) lacking E6AP escape replicative senescence, as well as Ras-induced senescence associated with impaired markers. E6AP-deficient MEFs exhibit a range of transformed phenotypes: these include the ability to grow under stress conditions (such as low serum and DNA damage), enhanced proliferation, anchorage independent growth and enhanced growth of xenografts in mice. The transformed phenotype of E6AP-deficient MEFs is associated with lower basal and stress-induced accumulation of p53. Overall, our study implicates E6AP as an important regulator of the cellular response to stress, in particular through the regulation of replicative and oncogene-induced senescence.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bardeesy N, Sinha M, Hezel AF, Signoretti S, Hathaway NA, Sharpless NE et al. (2002). Loss of the Lkb1 tumour suppressor provokes intestinal polyposis but resistance to transformation. Nature 419: 162–167.
Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N et al. (2006). Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444: 633–637.
Beaudenon S, Huibregtse JM . (2008). HPV E6, E6AP and cervical cancer. BMC Biochem 9 (Suppl 1): S4.
Blasco MA, Lee HW, Hande MP, Samper E, Lansdorp PM, DePinho RA et al. (1997). Telomere shortening and tumor formation by mouse cells lacking telomerase RNA. Cell 91: 25–34.
Campisi J . (2005). Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors. Cell 120: 513–522.
Campisi J, d'Adda di Fagagna F . (2007). Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 8: 729–740.
Ciechanover A . (2006). The ubiquitin proteolytic system: from a vague idea, through basic mechanisms, and onto human diseases and drug targeting. Neurology 66: S7–19.
Collado M, Serrano M . (2006). The power and the promise of oncogene-induced senescence markers. Nat Rev Cancer 6: 472–476.
Collado M, Serrano M . (2010). Senescence in tumours: evidence from mice and humans. Nat Rev Cancer 10: 51–57.
Dannenberg JH, van Rossum A, Schuijff Lte Riele H . (2000). Ablation of the retinoblastoma gene family deregulates G(1) control causing immortalization and increased cell turnover under growth-restricting conditions. Genes Dev 14: 3051–3064.
Di Micco R, Cicalese A, Fumagalli M, Dobreva M, Verrecchia A, Pelicci PG et al. (2008). DNA damage response activation in mouse embryonic fibroblasts undergoing replicative senescence and following spontaneous immortalization. Cell Cycle 7: 3601–3606.
Di Micco R, Fumagalli M, d'Adda di Fagagna F . (2007). Breaking news: high-speed race ends in arrest--how oncogenes induce senescence. Trends Cell Biol 17: 529–536.
Dimri GP . (2005). What has senescence got to do with cancer? Cancer Cell 7: 505–512.
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C et al. (1995). A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 92: 9363–9367.
Ferbeyre G, de Stanchina E, Lin AW, Querido E, McCurrach ME, Hannon GJ et al. (2002). Oncogenic ras and p53 cooperate to induce cellular senescence. Mol Cell Biol 22: 3497–3508.
Frank KM, Sharpless NE, Gao Y, Sekiguchi JM, Ferguson DO, Zhu C et al. (2000). DNA ligase IV deficiency in mice leads to defective neurogenesis and embryonic lethality via the p53 pathway. Mol Cell 5: 993–1002.
Gil J, Peters G . (2006). Regulation of the INK4b-ARF-INK4a tumour suppressor locus: all for one or one for all. Nat Rev Mol Cell Biol 7: 667–677.
Halazonetis TD, Gorgoulis VG, Bartek J . (2008). An oncogene-induced DNA damage model for cancer development. Science 319: 1352–1355.
Harvey M, McArthur MJ, Montgomery Jr CA, Bradley A, Donehower LA . (1993). Genetic background alters the spectrum of tumors that develop in p53-deficient mice. FASEB J 7: 938–943.
Haupt S, di Agostino S, Mizrahi I, Alsheich-Bartok O, Voorhoeve M, Damalas A et al. (2009). Promyelocytic leukemia protein is required for gain of function by mutant p53. Cancer Res 69: 4818–4826.
Haupt Y, Barak Y, Oren M . (1996). Cell type-specific inhibition of p53-mediated apoptosis by mdm2. EMBO J 15: 1596–1606.
Howley PM . (2006). Warts, cancer and ubiquitylation: lessons from the papillomaviruses. Trans Am Clin Climatol Assoc 117: 113–126; discussion 126-117.
Huibregtse JM, Scheffner M, Howley PM . (1991). A cellular protein mediates association of p53 with the E6 oncoprotein of human papillomavirus types 16 or 18. EMBO J 10: 4129–4135.
Jiang Y, Tsai TF, Bressler J, Beaudet AL . (1998a). Imprinting in Angelman and Prader-Willi syndromes. Curr Opin Genet Dev 8: 334–342.
Jiang YH, Armstrong D, Albrecht U, Atkins CM, Noebels JL, Eichele G et al. (1998b). Mutation of the Angelman ubiquitin ligase in mice causes increased cytoplasmic p53 and deficits of contextual learning and long-term potentiation. Neuron 21: 799–811.
Khan OY, Fu G, Ismail A, Srinivasan S, Cao X, Tu Y et al. (2006). Multifunction steroid receptor coactivator, E6-associated protein, is involved in development of the prostate gland. Mol Endocrinol 20: 544–559.
Kortlever RM, Higgins PJ, Bernards R . (2006). Plasminogen activator inhibitor-1 is a critical downstream target of p53 in the induction of replicative senescence. Nat Cell Biol 8: 877–884.
Kuilman T, Michaloglou C, Mooi WJ, Peeper DS . (2010). The essence of senescence. Genes Dev 24: 2463–2479.
Lahav G, Rosenfeld N, Sigal A, Geva-Zatorsky N, Levine AJ, Elowitz MB et al. (2004). Dynamics of the p53-Mdm2 feedback loop in individual cells. Nat Genet 36: 147–150.
Lev Bar-Or R, Maya R, Segel LA, Alon U, Levine AJ, Oren M . (2000). Generation of oscillations by the p53-Mdm2 feedback loop: a theoretical and experimental study. Proc Natl Acad Sci USA 97: 11250–11255.
Longworth MS, Laimins LA . (2004). Pathogenesis of human papillomaviruses in differentiating epithelia. Microbiol Mol Biol Rev 68: 362–372.
Louria-Hayon I, Alsheich-Bartok O, Levav-Cohen Y, Silberman I, Berger M, Grossman T et al. (2009). E6AP promotes the degradation of the PML tumor suppressor. Cell Death Differ 16: 1156–1166.
Maezawa Y, Yokote K, Sonezaki K, Fujimoto M, Kobayashi K, Kawamura H et al. (2006). Influence of C-peptide on early glomerular changes in diabetic mice. Diabetes Metab Res Rev 22: 313–322.
Matentzoglu K, Scheffner M . (2008). Ubiquitin ligase E6-AP and its role in human disease. Biochem Soc Trans 36: 797–801.
Matsuura T, Sutcliffe JS, Fang P, Galjaard RJ, Jiang YH, Benton CS et al. (1997). De novo truncating mutations in E6-AP ubiquitin-protein ligase gene (UBE3A) in Angelman syndrome. Nat Genet 15: 74–77.
Mishra A, Godavarthi SK, Jana NR . (2009). UBE3A/E6-AP regulates cell proliferation by promoting proteasomal degradation of p27. Neurobiol Dis 36: 26–34.
Mishra A, Jana NR . (2008). Regulation of turnover of tumor suppressor p53 and cell growth by E6-AP, a ubiquitin protein ligase mutated in Angelman mental retardation syndrome. Cell Mol Life Sci 65: 656–666.
Mu XC, Higgins PJ . (1995). Differential growth state-dependent regulation of plasminogen activator inhibitor type-1 expression in senescent IMR-90 human diploid fibroblasts. J Cell Physiol 165: 647–657.
Papazoglu C, Mills AA . (2007). p53: at the crossroad between cancer and ageing. J Pathol 211: 124–133.
Parrinello S, Samper E, Krtolica A, Goldstein J, Melov S, Campisi J . (2003). Oxygen sensitivity severely limits the replicative lifespan of murine fibroblasts. Nat Cell Biol 5: 741–747.
Rodier F, Campisi J, Bhaumik D . (2007). Two faces of p53: aging and tumor suppression. Nucleic Acids Res 35: 7475–7484.
Rodier F, Coppe JP, Patil CK, Hoeijmakers WA, Munoz DP, Raza SR et al. (2009). Persistent DNA damage signallingtriggers senescence-associated inflammatory cytokine secretion. Nat Cell Biol 11: 973–979.
Sage J, Miller AL, Perez-Mancera PA, Wysocki JM, Jacks T . (2003). Acute mutation of retinoblastoma gene function is sufficient for cell cycle re-entry. Nature 424: 223–228.
Sage J, Mulligan GJ, Attardi LD, Miller A, Chen S, Williams B et al. (2000). Targeted disruption of the three Rb-related genes leads to loss of G(1) control and immortalization. Genes Dev 14: 3037–3050.
Scheffner M, Huibregtse JM, Vierstra RD, Howley PM . (1993). The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53. Cell 75: 495–505.
Scheffner M, Werness BA, Huibregtse JM, Levine AJ, Howley PM . (1990). The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell 63: 1129–1136.
Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW . (1997). Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88: 593–602.
Sherman MY, Gabai V, O'Callaghan C, Yaglom J . (2007). Molecular chaperones regulate p53 and suppress senescence programs. FEBS Lett 581: 3711–3715.
Sherr CJ, DePinho RA . (2000). Cellular senescence: mitotic clock or culture shock? Cell 102: 407–410.
Smith JR, Lincoln II DW . (1984). Aging of cells in culture. Int Rev Cytol 89: 151–177.
Talis AL, Huibregtse JM, Howley PM . (1998). The role of E6AP in the regulation of p53 protein levels in human papillomavirus (HPV)-positive and HPV-negative cells. J Biol Chem 273: 6439–6445.
Todaro GJ, Green H . (1963). Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol 17: 299–313.
Ventura A, Kirsch DG, McLaughlin ME, Tuveson DA, Grimm J, Lintault L et al. (2007). Restoration of p53 function leads to tumour regression in vivo. Nature 445: 661–665.
Wolyniec K, Wotton S, Kilbey A, Jenkins A, Terry A, Peters G et al. (2009). RUNX1 and its fusion oncoprotein derivative, RUNX1-ETO, induce senescence-like growth arrest independently of replicative stress. Oncogene 28: 2502–2512.
Zuckerman V, Wolyniec K, Sionov RV, Haupt S, Haupt Y . (2009). Tumour suppression by p53: the importance of apoptosis and cellular senescence. J Pathol 219: 3–15.
Zufferey R, Dull T, Mandel RJ, Bukovsky A, Quiroz D, Naldini L et al. (1998). Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 72: 9873–9880.
Acknowledgements
We express our thanks to Scott Lowe and Martin Scheffner for the generous gifts of expression plasmids, and to Mariam Mansour for critical comments on the manuscript. Work in the authors laboratory is supported by an NHMRC project grant (grant No. 509196), by a grant from the Cancer Council Victoria, by the VESKI award and by the EC FP6 funding of the European commission (contract 503576). This publication reflects only the authors' views. The European commission is not liable for any use that may be made of the information herein.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Levav-Cohen, Y., Wolyniec, K., Alsheich-Bartok, O. et al. E6AP is required for replicative and oncogene-induced senescence in mouse embryo fibroblasts. Oncogene 31, 2199–2209 (2012). https://doi.org/10.1038/onc.2011.402
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2011.402
Keywords
This article is cited by
-
Induction of E6AP by microRNA-302c dysregulation inhibits TGF-β-dependent fibrogenesis in hepatic stellate cells
Scientific Reports (2020)
-
Identifying the ubiquitination targets of E6AP by orthogonal ubiquitin transfer
Nature Communications (2017)
-
Restoration of tumor suppression in prostate cancer by targeting the E3 ligase E6AP
Oncogene (2016)
-
The E6AP E3 ubiquitin ligase regulates the cellular response to oxidative stress
Oncogene (2013)