Hypophosphorylation of Mdm2 Augments p53 Stability
Open accessAbstract
The Mdm2 protein mediates ubiquitylation and degradation of p53 and is a key regulator of this tumor suppressor. More recently, it has been shown that Mdm2 is highly phosphorylated within its central acidic domain. In order to address the issue of how these modifications might regulate Mdm2 function, putative phosphorylation sites within this domain were substituted, individually or in pairs, with alanine residues. Mutants with serine-to-alanine substitutions between residues 244 and 260 abolished or at least reduced the capacity of Mdm2 to promote p53 degradation. In each case, loss of degradation function was independent of the ability to bind to p53 or p14ARF. Moreover, each of the Mdm2 mutants completely retained the capacity to act as a ubiquitin ligase in vivo. Thus, ubiquitylation and degradation can be uncoupled. Two-dimensional phosphopeptide mapping coupled with the use of phospho-specific antibodies revealed that Mdm2 is phosphorylated physiologically at several sites within this region, consistent with the idea that phosphorylation is important for Mdm2 activity. Strikingly, treatment of cells with ionizing radiation resulted in a significant decrease in the phosphorylation of residues that are important for p53 turnover. This hypophosphorylation preceded p53 accumulation. These findings indicate that Mdm2 contributes an additional function toward the degradation of p53 that is distinct from its ubiquitin ligase activity and is regulated by phosphorylation. Our model suggests that hypophosphorylation of Mdm2 in response to ionizing irradiation inactivates this novel function, thereby contributing to p53 stabilization.
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Both endogenous Mdm2 and transfected Mdm2 were exclusively located in the nucleus independent of phosphorylation status of Mdm2.
“both endogenous Mdm2 (Fig. 5A and Fig. 6, B and C) and transfected Mdm2 (31,51,60) were exclusively located in the nucleus independent of phosphorylation status of Mdm2.”
From Stabilization of Mdm2 via Decreased Ubiquitination Is Mediated by Protein Kinase B/Akt-dependent Phosphorylation (2004) · cites this paper as [51]
Cites
High-Efficiency Transformation of Mammalian Cells by Plasmid DNA
Chen, C., & Okayama, H. (1987) · Molecular and Cellular Biology · doi:10.1128/mcb.7.8.2745 · Cited by 5,374 · see citing works
Mdm2 promotes the rapid degradation of p53
Haupt, Y., Maya, R., Kazaz, A., & Oren, M. (1997) · Nature · doi:10.1038/387296a0 · Cited by 4,506 · see citing works
Regulation of p53 stability by Mdm2
Kubbutat, M. H. G., Jones, S. N., & Vousden, K. H. (1997) · Nature · doi:10.1038/387299a0 · Cited by 3,312 · see citing works
Momand, J., Zambetti, G. P., Olson, D. C., George, D., & Levine, A. J. (1992) · Cell · doi:10.1016/0092-8674(92)90644-r · Cited by 3,113 · see citing works
Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53
Honda, R., Tanaka, H., & Yasuda, H. (1997) · FEBS Letters · doi:10.1016/s0014-5793(97)01480-4 · Cited by 1,951 · see citing works
Pomerantz, J., Schreiber-Agus, N., Liégeois, N. J., Silverman, A., Alland, L., Chin, L., Potes, J., Chen, K., Orlow, I., Lee, H.-W., et al. (1998) · Cell · doi:10.1016/s0092-8674(00)81400-2 · Cited by 1,521 · see citing works
Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53
Oliner, J. D., Pietenpol, J. A., Thiagalingam, S., Gyuris, J., Kinzler, K. W., & Vogelstein, B. (1993) · Nature · doi:10.1038/362857a0 · Cited by 1,447 · see citing works
p53: A Frequent Target for Genetic Abnormalities in Lung Cancer
Takahashi, T., Nau, M. M., Chiba, I., Birrer, M. J., Rosenberg, R. K., Vinocour, M., Levitt, M. L., Pass, H. I., Gazdar, A. F., & Minna, J. D. (1989) · Science · doi:10.1126/science.2554494 · Cited by 1,196 · see citing works
Stott, F. J., Bates, S., James, M. C., McConnell, B. B., Starborg, M., Brookes, S., Palmero, I., Ryan, K. M., Hara, E., Vousden, K. H., et al. (1998) · The EMBO Journal · doi:10.1093/emboj/17.17.5001 · Cited by 1,165 · see citing works
Nucleolar Arf sequesters Mdm2 and activates p53
Weber, J. D., Taylor, L. J., Roussel, M. F., Sherr, C. J., & Bar-Sagi, D. (1999) · Nature Cell Biology · doi:10.1038/8991 · Cited by 940 · see citing works
Ubiquitin-dependent c-Jun degradation in vivo is mediated by the δ domain
Treier, M., Staszewski, L. M., & Bohmann, D. (1994) · Cell · doi:10.1016/s0092-8674(94)90502-9 · Cited by 928 · see citing works
Association of p19ARF with Mdm2 inhibits ubiquitin ligase activity of Mdm2 for tumor suppressor p53
Honda, R., & Yasuda, H. (1999) · The EMBO Journal · doi:10.1093/emboj/18.1.22 · Cited by 724 · see citing works
Waseem, N. H., & Lane, D. P. (1990) · Journal of Cell Science · doi:10.1242/jcs.96.1.121 · Cited by 722 · see citing works
Self-citationATM-dependent phosphorylation of Mdm2 on serine 395: role in p53 activation by DNA damage
Maya, R., Balass, M., Kim, S.-T., Shkedy, D., Leal, J.-F. M., Shifman, O., Moas, M., Buschmann, T., Ronai, Z., Shiloh, Y., et al. (2001) · Genes & Development · doi:10.1101/gad.886901 · Cited by 716 · see citing works
Mapping of the p53 and mdm-2 Interaction Domains
Chen, J., Maréchal, V., & Levine, A. J. (1993) · Molecular and Cellular Biology · doi:10.1128/mcb.13.7.4107 · Cited by 689 · see citing works
Twenty years of p53 research: structural and functional aspects of the p53 protein
May, P., & May, E. (1999) · Oncogene · doi:10.1038/sj.onc.1203285 · Cited by 663 · see citing works
MDM2 — master regulator of the p53 tumor suppressor protein
Momand, J., Wu, H.-H., & Dasgupta, G. (2000) · Gene · doi:10.1016/s0378-1119(99)00487-4 · Cited by 641 · see citing works
The cellular response to p53: the decision between life and death
Sionov, R. V., & Haupt, Y. (1999) · Oncogene · doi:10.1038/sj.onc.1203130 · Cited by 600 · see citing works
Regulation of the p53 Tumor Suppressor Protein
Oren, M. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.51.36031 · Cited by 561 · see citing works
Rapid ATM-dependent phosphorylation of MDM2 precedes p53 accumulation in response to DNA damage
Khosravi, R., Maya, R., Gottlieb, T., Oren, M., Shiloh, Y., & Shkedy, D. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.26.14973 · Cited by 461 · see citing works
p300/MDM2 complexes participate in MDM2-mediated p53 degradation
Grossman, S. R., Perez, M., Kung, A. L., Joseph, M., Mansur, C., Xiao, Z.-X., Kumar, S., Howley, P. M., & Livingston, D. M. (1998) · Molecular Cell · doi:10.1016/s1097-2765(00)80140-9 · Cited by 414 · see citing works
Multiple C-Terminal Lysine Residues Target p53 for Ubiquitin-Proteasome-Mediated Degradation
Rodríguez, M. S., Desterro, J. M. P., Laı́n, S., Lane, D. P., & Hay, R. T. (2000) · Molecular and Cellular Biology · doi:10.1128/mcb.20.22.8458-8467.2000 · Cited by 386 · see citing works
Self-citationNishimura, H., Yerkes, E. B., Hohenfellner, K., et al. (1999) · Molecular Cell · doi:10.1016/s1097-2765(00)80169-0 · Cited by 368 · see citing works
Design of a synthetic Mdm2-binding mini protein that activates the p53 response in vivo
Böttger, A., Böttger, V., Sparks, A., Liu, W.-L., Howard, S. F., & Lane, D. P. (1997) · Current Biology · doi:10.1016/s0960-9822(06)00374-5 · Cited by 363 · see citing works
Self-citationCAHILLY, L. A., Yang‐Feng, T. L., Francke, U., & George, D. L. (1987) · Somatic Cell and Molecular Genetics · doi:10.1007/bf01535205 · Cited by 355 · see citing works
Midgley, C., Fisher, C., Bártek, J., Vojtěšek, B., Lane, D. P., & Barnes, D. M. (1992) · Journal of Cell Science · doi:10.1242/jcs.101.1.183 · Cited by 330 · see citing works
Self-citationBarak, Y., Gottlieb, E., Juven-Gershon, T., & Oren, M. (1994) · Genes & Development · doi:10.1101/gad.8.15.1739 · Cited by 328 · see citing works
Ubiquitination of p53 and p21 Is Differentially Affected by Ionizing and UV Radiation
Maki, C. G., & Howley, P. M. (1997) · Molecular and Cellular Biology · doi:10.1128/mcb.17.1.355 · Cited by 305 · see citing works
The ubiquitin-mediated proteolytic pathway: Mode of action and clinical implications
Ciechanover, A. J., Orian, A., & Schwartz, A. L. (2000) · Journal of Cellular Biochemistry · doi:10.1002/(sici)1097-4644(2000)77:34+ · Cited by 281 · see citing works
RB Regulates the Stability and the Apoptotic Function of p53 via MDM2
Hsieh, J.-K., Chan, F., O’Connor, D. J., Mittnacht, S., Zhong, S., & Lü, X. (1999) · Molecular Cell · doi:10.1016/s1097-2765(00)80309-3 · Cited by 255 · see citing works
Midgley, C., Desterro, J. M. P., Saville, M. K., Howard, S. F., Sparks, A., Hay, R. T., & Lane, D. P. (2000) · Oncogene · doi:10.1038/sj.onc.1203593 · Cited by 250 · see citing works
Self-citationBlattner, C., Sparks, A., & Lane, D. P. (1999) · Molecular and Cellular Biology · doi:10.1128/mcb.19.5.3704 · Cited by 213 · see citing works
Self-citationThe role of p53 in tumour suppression: lessons from mouse models
Attardi, L. D., & Jacks, T. (1999) · Cellular and Molecular Life Sciences · doi:10.1007/s000180050269 · Cited by 200 · see citing works
van der Geer, P., & Hunter, T. R. (1994) · Electrophoresis · doi:10.1002/elps.1150150173 · Cited by 137 · see citing works
The contribution of the acidic domain of MDM2 to p53 and MDM2 stability
Argentini, M., Barboule, N., & Wasylyk, B. (2001) · Oncogene · doi:10.1038/sj.onc.1204241 · Cited by 90 · see citing works
The Li-Fraumeni syndrome: An inherited susceptibility to cancer
Evans, S. C., & Lozano, G. (1997) · Molecular Medicine Today · doi:10.1016/s1357-4310(97)01105-2 · Cited by 82 · see citing works
Vojtěšek, B., Dolezalova, H., Lauerová, L., Svitáková, M., Havlis, P., Kovařík, J., Midgley, C. A., & Lane, D. P. (1995) · PubMed · Cited by 72 · see citing works
Self-citationMechanisms of ubiquitin-mediated, limited processingof the NF-κB1 precursor protein p105
Ciechanover, A. J., Gonen, H., Bercovich, B., Cohen, S., Fajerman, I., Israël, A., Mercurio, F., Kahana, C., Schwartz, A. L., Iwaï, K., et al. (2001) · Biochimie · doi:10.1016/s0300-9084(01)01239-1 · Cited by 63 · see citing works
Hay, T., & Meek, D. W. (2000) · FEBS Letters · doi:10.1016/s0014-5793(00)01850-0 · Cited by 61 · see citing works
Self-citationLeng, R. P., Brown, D. R., Shivakumar, C., Deb, S., & Deb, S. (1995) · PubMed · Cited by 48 · see citing works
Hjerrild, M., Milne, D., Dumaz, N., Hay, T., Issinger, O., & Meek, D. W. (2001) · Biochemical Journal · doi:10.1042/0264-6021:3550347 · Cited by 45 · see citing works
Self-citationHengstermann, A., Whitaker, N. J., Zimmer, D., Zentgraf, H., & Scheffner, M. (1998) · Oncogene · doi:10.1038/sj.onc.1202282 · Cited by 32 · see citing works
Hjerrild, M., MILNE, D. M., Dumaz, N., et al. (2001) · Biochemical Journal · doi:10.1042/bj3550347 · Cited by 26 · see citing works
Meek, D. W., & MILNE, D. M. (2003) · Humana Press eBooks · doi:10.1385/1-59259-054-3:447 · Cited by 9 · see citing works
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Post-translational modification of p53 in tumorigenesis
Bode, A. M., & Dong, Z. (2004) · Nature Reviews Cancer · doi:10.1038/nrc1455 · Cited by 1,270 · see citing works
p53 ubiquitination: Mdm2 and beyond
Brooks, C. L., & Gu, W. (2006) · Molecular Cell · doi:10.1016/j.molcel.2006.01.020 · Cited by 924 · see citing works
Mono- Versus Polyubiquitination: Differential Control of p53 Fate by Mdm2
Li, M., Brooks, C. L., Wu-Baer, F., Chen, D., Baer, R., & Gu, W. (2003) · Science · doi:10.1126/science.1091362 · Cited by 804 · see citing works
Feng, J., Tamaskovic, R., Yang, Z., Brazil, D. P., Merlo, A., Hess, D., & Hemmings, B. A. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m404936200 · Cited by 294 · see citing works
Glycogen synthase kinase 3-dependent phosphorylation of Mdm2 regulates p53 abundance
Kulikov, R., Boehme, K. A., & Blattner, C. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.25.16.7170-7180.2005 · Cited by 132 · see citing works
Self-citationCK1α Plays a Central Role in Mediating MDM2 Control of p53 and E2F-1 Protein Stability
Huart, A.-S., MacLaine, N. J., Meek, D. W., & Hupp, T. R. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.m109.052647 · Cited by 110 · see citing works
Self-citationMdm2 facilitates the association of p53 with the proteasome
Kulikov, R., Letienne, J., Kaur, M., Grossman, S. R., Arts, J., & Blattner, C. (2010) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0911716107 · Cited by 94 · see citing works
Self-citationFunctional role of Mdm2 phosphorylation by ATR in attenuation of p53 nuclear export
Shinozaki, T., Nota, A., Taya, Y., & Okamoto, K. (2003) · Oncogene · doi:10.1038/sj.onc.1207176 · Cited by 93 · see citing works
Regulation of p53-MDMX interaction by casein kinase 1 alpha
Chen, L., Li, C., Pan, Y., & Chen, J. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.25.15.6509-6520.2005 · Cited by 88 · see citing works
Meek, D. W., & Hupp, T. R. (2010) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2009.10.005 · Cited by 76 · see citing works
Self-citationWinter, M., Milne, D., Dias, S., Kulikov, R., Knippschild, U., Blattner, C., & Meek, D. (2004) · Biochemistry · doi:10.1021/bi0489255 · Cited by 60 · see citing works
The roles of MDM2 and MDMX phosphorylation in stress signaling to p53
Chen, J. (2012) · Genes & Cancer · doi:10.1177/1947601912454733 · Cited by 33 · see citing works
The phenotype of MDM2 auto-degradation after DNA damage is due to epitope masking by phosphorylation
Cheng, Q., & Chen, J. (2011) · Cell Cycle · doi:10.4161/cc.10.7.15249 · Cited by 21 · see citing works
- Authors
- Blattner, C., Hay, T., Meek, D. W., & Lane, D. P.
- Affiliations
- Forschungszentrum Karlsruhe, Institute of Genetics & Toxicology, 76021 Karlsruhe, Germany
- Cancer Research Campaign Cell Transformation Group, Department of Surgery & Molecular Oncology
- University of Dundee (GB)
- Corresponding author
- Blattner, Christine
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2002-09-01
- Volume
- 22
- Issue
- 17
- Pages
- 6170-6182
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC134018
- OpenAlex
- W2134507382
- Cited by
- 144
- References
- 44
- Citations by year
- 2026: 2 · 2024: 1 · 2022: 3 · 2021: 1 · 2020: 1 · 2019: 1 · 2018: 2 · 2017: 3 · 2016: 6 · 2015: 5 · 2014: 4 · 2013: 5 · 2012: 8
- FWCI
- 6.67
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular and Cellular Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Discovery Research Portal (University of Dundee) (submittedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Medical Research Council
- Topics
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Keywords
- MDM2, p53, hypophosphorylation, p53 stabilization, p53 degradation, ubiquitination, ubiquitin ligase, acidic domain, serine phosphorylation, ionizing radiation, p14ARF, tumor suppressor regulation
- MeSH terms
- Amino Acid Sequence, Animals, Binding Sites, Carcinoma, Cells, Cultured, Chlorocebus aethiops, Cysteine Endopeptidases, Fibroblasts, Gamma Rays, Humans, Lung Neoplasms, Molecular Sequence Data, Multienzyme Complexes, Nuclear Proteins, Peptide Mapping, Phosphorylation, Protein Processing, Post-Translational, Proto-Oncogene Proteins, Recombinant Fusion Proteins, Species Specificity, Transfection, Tumor Cells, Cultured, Tumor Suppressor Protein p53, Mutagenesis, Site-Directed, Sequence Deletion, Protein Structure, Tertiary, COS Cells, Ubiquitin, Tumor Suppressor Protein p14ARF, Proteasome Endopeptidase Complex, Mice, Proto-Oncogene Proteins c-mdm2
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