The phenotype of MDM2 auto-degradation after DNA damage is due to epitope masking by phosphorylation
Open accessAbstract
It is widely accepted that DNA damage induces rapid degradation of MDM2 through phosphorylation, resulting in a transient reduction of MDM2 level. Elimination of MDM2 is a logical mechanism that stabilizes p53. This phenomenon has been reproduced by many independent studies and is frequently referenced. Here we present evidence that only phosphorylation-sensitive antibodies SMP14 and 2A10, but not other MDM2 antibodies, can detect robust down-regulation of MDM2 after DNA damage. Therefore, we conclude that DNA damage does not accelerate MDM2 auto-degradation. SMP14 and 2A10 are frequently used to detect human and mouse MDM2, respectively. While it is not clear whether the discrepancy is entirely due to the use of these antibodies, our results suggest that epitope masking by phosphorylation should be an important consideration when interpreting results of MDM2 analysis by SMP14 and 2A10.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The 2A9 MDM2 antibody was used to avoid possible epitope masking due to MDM2 phosphorylation.
“The 2A9 MDM2 antibody was used to avoid possible epitope masking due to phosphorylation of MDM2.11”
From Combination therapy in a xenograft model of glioblastoma: Enhancement of the antitumor activity of temozolomide by an MDM2 antagonist (2017) · cites this paper as [11]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
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
DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2
Shieh, S.-Y., Ikeda, M., Taya, Y., & Prives, C. (1997) · Cell · doi:10.1016/s0092-8674(00)80416-x · Cited by 2,100 · see citing works
The p53 pathway: positive and negative feedback loops
Harris, S. L., & Levine, A. J. (2005) · Oncogene · doi:10.1038/sj.onc.1208615 · Cited by 1,967 · 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
The p53-mdm-2 autoregulatory feedback loop.
Wu, X., Bayle, J. H., Olson, D., & Levine, A. J. (1993) · Genes & Development · doi:10.1101/gad.7.7a.1126 · Cited by 1,801 · see citing works
Enhanced Phosphorylation of p53 by ATM in Response to DNA Damage
Banin, S., Moyal, L., Shieh, S.-Y., Taya, Y., Anderson, C. W., Chessa, L., Smorodinsky, N. I., Prives, C., Reiss, Y., Shiloh, Y., et al. (1998) · Science · doi:10.1126/science.281.5383.1674 · Cited by 1,797 · see citing works
Rescue of early embryonic lethality in mdm2-deficient mice by deletion of p53
de Oca Luna, R. M., Wagner, D. S., & Lozano, G. (1995) · Nature · doi:10.1038/378203a0 · Cited by 1,420 · cited 1× · see citing works
“However, current evidence suggests that MDM2 is a major and indispensable regulator of p53 level.(7,8) Elucidating how p53 is stabilized and activated after DNA damage is the subject of many studies.”
Backgroundmdm2 expression is induced by wild type p53 activity.
Barak, Y., Juven, T., Haffner, R., & Oren, M. (1993) · The EMBO Journal · doi:10.1002/j.1460-2075.1993.tb05678.x · Cited by 1,290 · see citing works
Rescue of embryonic lethality in Mdm2-deficient mice by absence of p53
Jones, S. N., Roe, A. E., Donehower, L. A., & Bradley, A. (1995) · Nature · doi:10.1038/378206a0 · Cited by 1,280 · cited 1× · see citing works
“However, current evidence suggests that MDM2 is a major and indispensable regulator of p53 level.(7,8) Elucidating how p53 is stabilized and activated after DNA damage is the subject of many studies.”
BackgroundMayo, L. D., & Donner, D. B. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.181181198 · Cited by 1,230 · see citing works
Shieh, S.-Y., Ahn, J., Tamai, K., Taya, Y., & Prives, C. (2000) · Genes & Development · doi:10.1101/gad.14.3.289 · Cited by 972 · see citing works
Chk2/hCds1 functions as a DNA damage checkpoint in G<sub>1</sub>by stabilizing p53
Chehab, N. H., Malikzay, A., Appel, M., & Halazonetis, T. D. (2000) · Genes & Development · doi:10.1101/gad.14.3.278 · Cited by 805 · see citing works
Pirh2, a p53-Induced Ubiquitin-Protein Ligase, Promotes p53 Degradation
Leng, R. P., Lin, Y., Ma, W., Wu, H., Lemmers, B., Chung, S., Parant, J. M., Lozano, G., Hakem, R., & Benchimol, S. (2003) · Cell · doi:10.1016/s0092-8674(03)00193-4 · Cited by 717 · see citing works
ATM-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
The ubiquitin ligase COP1 is a critical negative regulator of p53
Dornan, D., Wertz, I. E., Shimizu, H., Arnott, D. P., Frantz, G. D., Dowd, P., Rourke, K. O., Koeppen, H., & Dixit, V. M. (2004) · Nature · doi:10.1038/nature02514 · Cited by 700 · 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
Self-citationRapid 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
Accelerated MDM2 auto‐degradation induced by DNA‐damage kinases is required for p53 activation
Stommel, J. M., & Wahl, G. M. (2004) · The EMBO Journal · doi:10.1038/sj.emboj.7600145 · Cited by 386 · see citing works
ATM and Chk2-dependent phosphorylation of MDMX contribute to p53 activation after DNA damage
Chen, L., Gilkes, D. M., Pan, Y., Lane, W. S., & Chen, J. (2005) · The EMBO Journal · doi:10.1038/sj.emboj.7600812 · Cited by 260 · see citing works
Self-citation(1997) · PubMed · Cited by 243 · see citing works
Picksley, S. M., Vojtĕsek, B., Sparks, A., & Lane, D. P. (1994) · PubMed · Cited by 238 · see citing works
Inuzuka, H., Tseng, A., Gao, D., Zhai, B., Zhang, Q. H., Shaik, S., Wan, L., Cambronne, X. A., Mock, C. S., Yin, H., et al. (2010) · Cancer Cell · doi:10.1016/j.ccr.2010.06.015 · Cited by 205 · see citing works
Phosphorylation of HDM2 by Akt
Ashcroft, M., Ludwig, R. L., Woods, D. B., Copeland, T. D., Weber, H. O., MacRae, E. J., & Vousden, K. H. (2002) · Oncogene · doi:10.1038/sj.onc.1205276 · Cited by 197 · see citing works
ATM activates p53 by regulating MDM2 oligomerization and E3 processivity
Cheng, Q., Chen, L., Li, Z., Lane, W. S., & Chen, J. (2009) · The EMBO Journal · doi:10.1038/emboj.2009.294 · Cited by 165 · see citing works
Self-citationSer18 and 23 phosphorylation is required for p53‐dependent apoptosis and tumor suppression
Chao, C., Herr, D., Chun, J., & Xu, Y. (2006) · The EMBO Journal · doi:10.1038/sj.emboj.7601167 · Cited by 158 · see citing works
Hypophosphorylation of Mdm2 Augments p53 Stability
Blattner, C., Hay, T., Meek, D. W., & Lane, D. P. (2002) · Molecular and Cellular Biology · doi:10.1128/mcb.22.17.6170-6182.2002 · Cited by 144 · see citing works
Cell Type- and Promoter-specific Roles of Ser18 Phosphorylation in Regulating p53 Responses
Chao, C., Hergenhahn, M., Kaeser, M. D., Wu, Z., Saito, S., Iggo, R., Hollstein, M., Appella, E., & Xu, Y. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m306938200 · Cited by 141 · see citing works
Defective apoptosis and B-cell lymphomas in mice with p53 point mutation at Ser 23
MacPherson, D., Kim, J., Kim, T., Rhee, B. K., van Oostrom, C. T., DiTullio, R. A., Venere, M., Halazonetis, T. D., Bronson, R., de Vries, A., et al. (2004) · The EMBO Journal · doi:10.1038/sj.emboj.7600363 · Cited by 124 · see citing works
Vogt Sionov, R., Coen, S., Goldberg, Z., Berger, M., Bercovich, B., Ben-Neriah, Y., Ciechanover, A., & Haupt, Y. (2001) · Molecular and Cellular Biology · doi:10.1128/mcb.21.17.5869-5878.2001 · Cited by 94 · see citing works
Functional 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
Binding of p53 to the Central Domain of Mdm2 Is Regulated by Phosphorylation
Kulikov, R., Winter, M., & Blattner, C. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m513311200 · Cited by 79 · see citing works
Cyclin A-CDK Phosphorylation Regulates MDM2 Protein Interactions
Zhang, T., & Prives, C. L. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m011326200 · Cited by 70 · see citing works
de Toledo, S. M., Azzam, E. I., Dahlberg, W. K., Gooding, T. B., & Little, J. B. (2000) · Oncogene · doi:10.1038/sj.onc.1204020 · Cited by 64 · see citing works
Maya, R., & Oren, M. (2000) · Oncogene · doi:10.1038/sj.onc.1203658 · Cited by 41 · see citing works
Flagged first, then by citations
1 of the 37 references couldn’t be matched to a known paper.
Research works printed but not linked: 1
- Prives C, Hall PA. The p53 pathway. J Pathol 1999; 187:112-26.
Cited by
Sorted by citations
The role of ubiquitin modification in the regulation of p53
Hock, A. K., & Vousden, K. H. (2014) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/j.bbamcr.2013.05.022 · Cited by 168 · see citing works
Pant, V., Xiong, S., Iwakuma, T., Quintás-Cardama, A., & Lozano, G. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1102241108 · Cited by 134 · see citing works
SCF ubiquitin ligases in the maintenance of genome stability
Silverman, J. S., Skaar, J. R., & Pagano, M. (2011) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2011.10.004 · Cited by 89 · see citing works
The Mdm network and its regulation of p53 activities: a rheostat of cancer risk
Eischen, C. M., & Lozano, G. (2014) · Human Mutation · doi:10.1002/humu.22524 · Cited by 79 · see citing works
Cullin Ring Ubiquitin Ligases (CRLs) in Cancer: Responses to Ionizing Radiation (IR) Treatment
Fouad, S., Wells, O. S., Hill, M. A., & D’Angiolella, V. (2019) · Frontiers in Physiology · doi:10.3389/fphys.2019.01144 · Cited by 63 · see citing works
Wang, H., Cai, S., Bailey, B. J., Reza Saadatzadeh, M., Ding, J., Tonsing-Carter, E., Georgiadis, T. M., Zachary Gunter, T., Long, E. C., Minto, R. E., et al. (2017) · Journal of Neurosurgery · doi:10.3171/2016.1.jns152513 · Cited by 49 · see citing works
SMG-1 suppresses CDK2 and tumor growth by regulating both the p53 and Cdc25A signaling pathways
Gubanova, E., Issaeva, N., Göktürk, C., Djureinovic, T., & Helleday, T. (2013) · Cell Cycle · doi:10.4161/cc.26660 · Cited by 41 · see citing works
FKBP12 enhances sensitivity to chemotherapy-induced cancer cell apoptosis by inhibiting MDM2
Liu, T., Xiong, J., Yi, S., Zhang, H., Zhou, S., Gu, L., & Zhou, M. L. (2016) · Oncogene · doi:10.1038/onc.2016.331 · Cited by 42 · see citing works
Structural basis for DNA damage-induced phosphoregulation of MDM2 RING domain
Magnussen, H., Ahmed, S. F., Sibbet, G. J., Hristova, V. A., Nomura, K., Hock, A. K., Archibald, L. J., Jamieson, A. G., Fushman, D., Vousden, K. H., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-15783-y · Cited by 37 · 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
Self-citationPham, T.-N. D., Ma, W., Miller, D., Kazakova, L., & Benchimol, S. (2019) · Cell Death and Disease · doi:10.1038/s41419-018-1274-6 · Cited by 27 · see citing works
A novel mechanism of crosstalk between the p53 and NFκB pathways
Heyne, K., Winter, C., Gerten, F., Schmidt, C., & Roemer, K. (2013) · Cell Cycle · doi:10.4161/cc.25495 · Cited by 22 · see citing works
Protecting the genome from mdm2 and mdmx
Melo, A. N., & Eischen, C. M. (2012) · Genes & Cancer · doi:10.1177/1947601912454139 · Cited by 20 · see citing works
Slabáková, E., Kharaishvili, G., Smějová, M., Pernicová, Z., Suchánková, T., Remšík, J., Lerch, S., Straková, N., Bouchal, J., Král, M., et al. (2015) · Oncotarget · doi:10.18632/oncotarget.5392 · Cited by 20 · see citing works
Transcriptional repressor NIR interacts with the p53-inhibiting ubiquitin ligase MDM2
Heyne, K., Förster, J., Schüle, R., & Roemer, K. (2014) · Nucleic Acids Research · doi:10.1093/nar/gkt1371 · Cited by 19 · see citing works
Senturk, J. C., Bohlman, S., & Manfredi, J. J. (2017) · Oncogene · doi:10.1038/onc.2017.229 · Cited by 18 · see citing works
Induction of ASAP (MAP9) contributes to p53 stabilization in response to DNA damage
Basbous, J., Knani, D., Bonneaud, N., Giorgi, D., Brondello, J., & Rouquier, S. (2012) · Cell Cycle · doi:10.4161/cc.20858 · Cited by 17 · see citing works
Development and characterization of 5 canine B-cell lymphoma cell lines
Zwingenberger, A. L., Vernau, W., Shi, C., Yan, W., Chen, X., Gordon, I. K., & Kent, M. S. (2011) · Leukemia Research · doi:10.1016/j.leukres.2011.11.004 · Cited by 6 · see citing works
MDM2 provides TOP2 poison resistance by promoting proteolysis of TOP2βcc in a p53-independent manner
Shu, J., Jiang, J., Wang, X., Yang, X., Zhao, G., & Cai, T. (2024) · Cell Death and Disease · doi:10.1038/s41419-024-06474-3 · Cited by 4 · see citing works
Lanicheva, A. K., Valeeva, R. R., & Semchenko, V. V. (2025) · Ulyanovsk Medico-biological Journal · doi:10.34014/2227-1848-2025-2-141-149
- Authors
- Cheng, Q., & Chen, J.
- Affiliations
- Moffitt Cancer Center (US)
- Venue
- Cell Cycle
- Publisher
- Informa UK Limited
- Published
- 2011-04
- Volume
- 10
- Issue
- 7
- Pages
- 1162-1166
- ISSN
- 1538-4101 (ISSN-L)
- 1551-4005
- PMC
- PMC3100889
- OpenAlex
- W1986441880
- Cited by
- 21
- References
- 37
- Citations by year
- 2025: 1 · 2024: 1 · 2020: 1 · 2019: 2 · 2017: 1 · 2016: 2 · 2015: 1 · 2014: 3 · 2013: 3 · 2012: 3
- FWCI
- 1.89
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Cell Cycle (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Cancer Institute (CA141244, R01CA141244)
- Topics
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- MDM2, protein degradation, DNA damage, phosphorylation, epitope masking, p53 stabilization, antibody specificity, p53 pathway
- MeSH terms
- Animals, Antibodies, Epitopes, Cell Line, DNA Damage, Gene Expression Regulation, Humans, Phosphorylation, Blotting, Western, Tumor Suppressor Protein p53, Mice, Proto-Oncogene Proteins c-mdm2
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
