Targeted inactivation of Mdm2 RING finger E3 ubiquitin ligase activity in the mouse reveals mechanistic insights into p53 regulation
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It is believed that Mdm2 suppresses p53 in two ways: transcriptional inhibition by direct binding, and degradation via its E3 ligase activity. To study these functions physiologically, we generated mice bearing a single-residue substitution (C462A) abolishing the E3 function without affecting p53 binding. Unexpectedly, homozygous mutant mice died before E7.5, and deletion of p53 rescued the lethality. Furthermore, reintroducing a switchable p53 by crossing with p53ER(TAM) mice surprisingly demonstrated that the mutant Mdm2(C462A) was rapidly degraded in a manner indistinguishable from that of the wild-type Mdm2. Hence, our data indicate that (1) the Mdm2-p53 physical interaction, without Mdm2-mediated p53 ubiquitination, cannot control p53 activity sufficiently to allow early mouse embryonic development, and (2) Mdm2's E3 function is not required for Mdm2 degradation.
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A knock-in mouse Mdm2 mutant with a C462A mutation in the RING domain was embryonic lethal in a p53-dependent manner, providing evidence that the Mdm2 RING domain is required for correct regulation of p53 activity in vivo.
“Recently, a knock-in mouse Mdm2 mutant was described in which one of the critical cysteines in the Mdm2 RING domain was mutated to alanine (C462A) [31]. The homozygous mutation was embryonic lethal in a p53-dependent manner, providing compelling evidence that the Mdm2 RING domain is required for correct regulation of p53 activity in vivo.”
From The p53 orchestra: Mdm2 and Mdmx set the tone (2010) · cites this paper as [31]
Loss of Mdm2 E3 ubiquitin ligase activity is likely responsible for the C462A phenotype, since the mutant could still bind p53 and block p53-dependent transactivation.
“The authors concluded that loss of Mdm2 E3 ubiquitin ligase activity is likely responsible for the observed phenotype, since the mutant could still bind to p53 and block p53-dependent transactivation.”
From The p53 orchestra: Mdm2 and Mdmx set the tone (2010) · cites this paper as [31]
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
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
p53: puzzle and paradigm.Preprint
Ko, L. J., & Prives, C. L. (1996) · Genes & Development · doi:10.1101/gad.10.9.1054 · Cited by 2,338 · 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
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
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 4× · see citing works
“Mice with targeted deletion of the Mdm2 gene die during early embryonic development, and this lethality can be rescued by concomitant deletion of p53, indicating that the embryonic lethality of Mdm2 null mice is due to activation of p53 (Jones et al., 1995; Luna et al., 1995).”
BackgroundInfluentialRescue 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 4× · see citing works
“Mice with targeted deletion of the Mdm2 gene die during early embryonic development, and this lethality can be rescued by concomitant deletion of p53, indicating that the embryonic lethality of Mdm2 null mice is due to activation of p53 (Jones et al., 1995; Luna et al., 1995).”
BackgroundInfluentialThe MDM2 gene amplification database
Momand, J. (1998) · Nucleic Acids Research · doi:10.1093/nar/26.15.3453 · Cited by 982 · see citing works
Michalovitz, D., Halevy, O., & Oren, M. (1990) · Cell · doi:10.1016/0092-8674(90)90113-s · Cited by 769 · see citing works
Lin, J., Chen, J., Elenbaas, B., & Levine, A. J. (1994) · Genes & Development · doi:10.1101/gad.8.10.1235 · Cited by 652 · see citing works
p53 Mutation and MDM2 amplification in human soft tissue sarcomas.
Leach, F. S., Tokino, T., Meltzer, P. S., Burrell, M., Oliner, J., Smith, S., HILL, D. E., Sidransky, D., Kinzler, K. W., & Vogelstein, B. (1993) · PubMed · Cited by 533 · see citing works
Itahana, K., Bhat, K., Jin, A., Itahana, Y., Hawke, D. H., Kobayashi, R., & Zhang, Y. (2003) · Molecular Cell · doi:10.1016/s1097-2765(03)00431-3 · Cited by 431 · see citing works
Self-citationp300/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
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
Honda, R., & Yasuda, H. (2000) · Oncogene · doi:10.1038/sj.onc.1203464 · Cited by 376 · see citing works
The MDM2 RING-finger domain is required to promote p53 nuclear export
Geyer, R. K., Yu, Z. K., & Maki, C. G. (2000) · Nature Cell Biology · doi:10.1038/35023507 · Cited by 361 · see citing works
MDM2 interacts with MDMX through their RING finger domains
Tanimura, S., Ohtsuka, S., Mitsui, K., Shirouzu, K., Yoshimura, A., & Ohtsubo, M. (1999) · FEBS Letters · doi:10.1016/s0014-5793(99)00254-9 · Cited by 324 · see citing works
Stabilization of the MDM2 Oncoprotein by Interaction with the Structurally Related MDMX Protein
Sharp, D. A., Kratowicz, S. A., Sank, M. J., & George, D. L. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.53.38189 · Cited by 286 · see citing works
Repression of p53-mediated transcription by MDM2: a dual mechanism
Thut, C. J., Goodrich, J. A., & Tjian, R. (1997) · Genes & Development · doi:10.1101/gad.11.15.1974 · Cited by 272 · see citing works
Mdm4 and Mdm2 cooperate to inhibit p53 activity in proliferating and quiescent cells <i>in vivo</i>
Francoz, S., Froment, P., Bogaerts, S., De Clercq, S., Maetens, M., Doumont, G., Bellefroid, E., & Marine, J.-C. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0508476103 · Cited by 260 · see citing works
Mdm2 is critically and continuously required to suppress lethal p53 activity in vivo
Ringshausen, I., O'Shea, C. C., Finch, A. J., Swigart, L. B., & Evan, G. I. (2006) · Cancer Cell · doi:10.1016/j.ccr.2006.10.010 · Cited by 256 · see citing works
Self-citationMdmX Protects p53 from Mdm2-Mediated Degradation
Jackson, M. W., & Berberich, S. J. (2000) · Molecular and Cellular Biology · doi:10.1128/mcb.20.3.1001-1007.2000 · Cited by 211 · cited 3× · see citing works
“The Mdm2 RING domain also mediates homodimerization with itself and heterodimerization with MDMX (Jackson and Berberich, 2000; Sharp et al., 1999; Stad et al., 2000; Tanimura et al., 1999), findings confirmed recently in a paper describing the NMR solution structure of the Mdm2 RING domain dimer…”
BackgroundThe RING domain of Mdm2 mediates histone ubiquitylation and transcriptional repression
Minsky, N., & Oren, M. (2004) · Molecular Cell · doi:10.1016/j.molcel.2004.10.016 · Cited by 204 · see citing works
Poyurovsky, M. V., Priest, C., Kentsis, A., Borden, K. L., Pan, Z., Pavletich, N., & Prives, C. (2007) · The EMBO Journal · doi:10.1038/sj.emboj.7601465 · Cited by 203 · see citing works
Toledo, F., Krummel, K. A., Lee, C. J., Liu, C.-W., Rodewald, L.-W., Tang, M., & Wahl, G. M. (2006) · Cancer Cell · doi:10.1016/j.ccr.2006.03.014 · Cited by 199 · cited 1× · see citing works
“This is consistent with studies by other mouse models showing that Mdm2 is responsible for p53 degradation in vivo (Francoz et al., 2006; Toledo et al., 2006).”
ResultIdentification of a cryptic nucleolar-localization signal in MDM2
Lohrum, M. A. E., Ashcroft, M. A., Kubbutat, M. H. G., & Vousden, K. H. (2000) · Nature Cell Biology · doi:10.1038/35004057 · Cited by 186 · see citing works
Temporal dissection of p53 function in vitro and in vivo
Christophorou, M. A., Martı́n-Zanca, D., Soucek, L., Lawlor, E. R., Swigart, L. B., Verschuren, E. W., & Evan, G. I. (2005) · Nature Genetics · doi:10.1038/ng1572 · Cited by 182 · see citing works
Self-citationIntrinsic ubiquitination activity of PCAF controls the stability of the oncoprotein Hdm2
Linares, L. K., Kiernan, R. E., Triboulet, R., Chable-Bessia, C., Latreille, D., Cuvier, O., Lacroix, M., Cam, L. L., Coux, O., & Benkirane, M. (2007) · Nature Cell Biology · doi:10.1038/ncb1545 · Cited by 180 · see citing works
Regulating the p53 system through ubiquitination
Yang, Y., Li, C.-C. H., & Weissman, A. M. (2004) · Oncogene · doi:10.1038/sj.onc.1207411 · Cited by 179 · see citing works
The MDM2 Oncoprotein Binds Specifically to RNA through its RING Finger Domain
Elenbaas, B., Dobbelstein, M., Roth, J. C., Shenk, T. E., & Levine, A. J. (1996) · Molecular Medicine · doi:10.1007/bf03401903 · Cited by 164 · see citing works
Lindström, M. S., Jin, A., Deisenroth, C., Wolf, G. W., & Zhang, Y. (2006) · Molecular and Cellular Biology · doi:10.1128/mcb.01307-06 · Cited by 149 · see citing works
Self-citationStad, R., Ramos, Y. F. M., Little, N., Grivell, S., Attema, J., van der Eb, A. J., & Jochemsen, A. G. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m003496200 · Cited by 145 · see citing works
Solution structure of the Hdm2 C2H2C4 RING, a domain critical for ubiquitination of p53
Kostic, M., Matt, T., Martinez-Yamout, M. A., Dyson, H. J., & Wright, P. E. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.08.027 · Cited by 128 · cited 2× · see citing works
“, 1999), findings confirmed recently in a paper describing the NMR solution structure of the Mdm2 RING domain dimer (Kostic et al., 2006).”
BackgroundCritical Contribution of the MDM2 Acidic Domain to p53 Ubiquitination
Kawai, H., Wiederschain, D., & Yuan, Z.-M. (2003) · Molecular and Cellular Biology · doi:10.1128/mcb.23.14.4939-4947.2003 · Cited by 122 · see citing works
Nucleotide Binding by the MDM2 RING Domain Facilitates Arf-Independent MDM2 Nucleolar Localization
Poyurovsky, M. V., Jacq, X., Ma, C., Karni-Schmidt, O., Parker, P. J., Chalfie, M., Manley, J. L., & Prives, C. L. (2003) · Molecular Cell · doi:10.1016/s1097-2765(03)00400-3 · Cited by 58 · see citing works
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Deshaies, R. J., & Joazeiro, C. A. P. (2009) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.78.101807.093809 · Cited by 2,679 · see citing works
Kruse, J.-P., & Gu, W. (2009) · Cell · doi:10.1016/j.cell.2009.04.050 · Cited by 1,610 · see citing works
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Wade, M., Li, Y.-C., & Wahl, G. M. (2013) · Nature Reviews Cancer · doi:10.1038/nrc3430 · Cited by 1,279 · see citing works
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Zhang, Y., & Lu, H. (2009) · Cancer Cell · doi:10.1016/j.ccr.2009.09.024 · Cited by 591 · see citing works
Self-citationChauhan, D., Tian, Z., Nicholson, B., Kumar, K. G. S., Zhou, B., Carrasco, R., McDermott, J. L., Leach, C. A., Fulcinniti, M., Kodrasov, M. P., et al. (2012) · Cancer Cell · doi:10.1016/j.ccr.2012.08.007 · Cited by 589 · see citing works
The p53 orchestra: Mdm2 and Mdmx set the tone
Wade, M., Wang, Y. V., & Wahl, G. M. (2010) · Trends in Cell Biology · doi:10.1016/j.tcb.2010.01.009 · Cited by 450 · see citing works
Mdm2-mediated ubiquitylation: p53 and beyond
Marine, J.-C., & Lozano, G. (2010) · Cell Death & Differentiation · doi:10.1038/cdd.2009.68 · Cited by 354 · see citing works
Trim24 targets endogenous p53 for degradation
Allton, K., Jain, A. K., Herz, H.-M., Tsai, W.-W., Jung, S. Y., Qin, J., Bergmann, A., Johnson, R. L., & Barton, M. C. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0813177106 · Cited by 292 · see citing works
Linke, K., Mace, P. D., Smith, C. A., Vaux, D. L., Silke, J., & Day, C. L. (2008) · Cell Death & Differentiation · doi:10.1038/sj.cdd.4402309 · Cited by 288 · see citing works
The p53 inhibitors MDM2/MDMX complex is required for control of p53 activity in vivo
Huang, L., Yan, Z., Liao, X., Li, Y., Yang, J., Wang, Z.-G., Zuo, Y., Kawai, H., Shadfan, M., Ganapathy, S., et al. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1102309108 · Cited by 193 · see citing works
Targeting Mdm2 and Mdmx in cancer therapy: better living through medicinal chemistry?
Wade, M., & Wahl, G. M. (2009) · Molecular Cancer Research · doi:10.1158/1541-7786.mcr-08-0423 · Cited by 170 · see citing works
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
Regulation of p53: a collaboration between Mdm2 and MdmX
Pei, D., Zhang, Y., & Zheng, J. (2012) · Oncotarget · doi:10.18632/oncotarget.443 · Cited by 140 · see citing works
Self-citationPant, 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
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Herman, A. G., Hayano, M., Poyurovsky, M. V., Shimada, K., Skouta, R., Prives, C., & Stockwell, B. R. (2011) · Cancer Discovery · doi:10.1158/2159-8290.cd-11-0104 · Cited by 100 · see citing works
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Allende-Vega, N., Sparks, A., Lane, D. P., & Saville, M. K. (2010) · Oncogene · doi:10.1038/onc.2009.330 · Cited by 100 · see citing works
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- Authors
- Itahana, K., Mao, H., Jin, A., Itahana, Y., Clegg, H. V., Lindström, M. S., Bhat, K. P., Godfrey, V. L., Evan, G. I., & Zhang, Y.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- The University of Texas MD Anderson Cancer Center (US)
- Corresponding author
- Zhang, Yanping
- Venue
- Cancer Cell
- Publisher
- Elsevier BV
- Published
- 2007-10
- Volume
- 12
- Issue
- 4
- Pages
- 355-366
- ISSN
- 1535-6108 (ISSN-L)
- OpenAlex
- W2014563476
- Cited by
- 245
- References
- 36
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 5 · 2023: 5 · 2022: 9 · 2021: 12 · 2020: 13 · 2019: 6 · 2018: 9 · 2017: 13 · 2016: 14 · 2015: 12 · 2014: 22 · 2013: 14 · 2012: 21
- FWCI
- 9.75
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cancer Cell (publishedVersion)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- National Institutes of Health
- National Cancer Institute (K01CA087580, R01CA100302)
- 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 › TGF-β signaling in diseases
- Keywords
- MDM2, p53, E3 ubiquitin ligase, ring finger, p53 degradation, p53-MDM2 interaction, transcriptional repression, embryonic lethality, ubiquitination
- MeSH terms
- Animals, Cells, Cultured, DNA Damage, Embryo, Mammalian, Fibroblasts, Gamma Rays, Genotype, Gestational Age, Homozygote, Mice, Inbred C57BL, Mutation, Phenotype, Transcription, Genetic, Down-Regulation, Tumor Suppressor Protein p53, Mutagenesis, Site-Directed, Protein Structure, Tertiary, Mice, Knockout, Gene Expression Regulation, Developmental, Amino Acid Substitution, Proteasome Endopeptidase Complex, Mice, Proto-Oncogene Proteins c-mdm2
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