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EMBO J. Oct 15, 1997; 16(20): 6217–6229.
PMCID: PMC1326306

Regulation of ES cell differentiation by functional and conformational modulation of p53.


Embryonic stem (ES) cell lines were used to examine the role of p53 during in vitro differentiation. Undifferentiated ES cells express high levels of p53 exclusively in the wild-type conformation, immunoprecipitable by monoclonal antibody PAb246, and p53 was found to be functionally active as determined by its ability to bind DNA specifically and to activate transcription of target genes. Differentiation in vitro resulted in a decrease in the levels of p53 and in a shift in its conformational status to the mutant form, detectable by monoclonal antibody PAb240, with a concomitant loss of functional activity. The presence of functional p53 in the undifferentiated ES cells renders them hypersensitive to UV irradiation, whereas the differentiated cells were resistant to UV treatment. ES cells lacking p53 exhibit enhanced proliferation in both the undifferentiated and differentiated state, and apoptosis accompanying differentiation was found to be reduced. Furthermore, wild-type ES cells undergoing apoptosis expressed functional p53. Expression of the temperature-sensitive p53val135 mutant in wild-type ES cells resulted in a reduction of apoptosis accompanying differentiation when it adopted a mutant conformation at 39 degrees C. These data demonstrate that functional inactivation of p53 allows differentiating cells to escape from apoptosis, and suggest that the conformational switch could regulate the inactivation process.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Attardi LD, Lowe SW, Brugarolas J, Jacks T. Transcriptional activation by p53, but not induction of the p21 gene, is essential for oncogene-mediated apoptosis. EMBO J. 1996 Jul 15;15(14):3693–3701. [PMC free article] [PubMed]
  • Banerjee D, Lenz HJ, Schnieders B, Manno DJ, Ju JF, Spears CP, Hochhauser D, Danenberg K, Danenberg P, Bertino JR. Transfection of wild-type but not mutant p53 induces early monocytic differentiation in HL60 cells and increases their sensitivity to stress. Cell Growth Differ. 1995 Nov;6(11):1405–1413. [PubMed]
  • Belloc F, Dumain P, Boisseau MR, Jalloustre C, Reiffers J, Bernard P, Lacombe F. A flow cytometric method using Hoechst 33342 and propidium iodide for simultaneous cell cycle analysis and apoptosis determination in unfixed cells. Cytometry. 1994 Sep 1;17(1):59–65. [PubMed]
  • Brenner L, Muñoz-Antonia T, Vellucci VF, Zhou ZL, Reiss M. Wild-type p53 tumor suppressor gene restores differentiation of human squamous carcinoma cells but not the response to transforming growth factor beta. Cell Growth Differ. 1993 Dec;4(12):993–1004. [PubMed]
  • Chen X, Ko LJ, Jayaraman L, Prives C. p53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumor cells. Genes Dev. 1996 Oct 1;10(19):2438–2451. [PubMed]
  • Montes de Oca Luna R, Wagner DS, Lozano G. Rescue of early embryonic lethality in mdm2-deficient mice by deletion of p53. Nature. 1995 Nov 9;378(6553):203–206. [PubMed]
  • Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery CA, Jr, Butel JS, Bradley A. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 1992 Mar 19;356(6366):215–221. [PubMed]
  • Dulić V, Kaufmann WK, Wilson SJ, Tlsty TD, Lees E, Harper JW, Elledge SJ, Reed SI. p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest. Cell. 1994 Mar 25;76(6):1013–1023. [PubMed]
  • Ehinger M, Nilsson E, Persson AM, Olsson I, Gullberg U. Involvement of the tumor suppressor gene p53 in tumor necrosis factor-induced differentiation of the leukemic cell line K562. Cell Growth Differ. 1995 Jan;6(1):9–17. [PubMed]
  • el-Deiry WS, Kern SE, Pietenpol JA, Kinzler KW, Vogelstein B. Definition of a consensus binding site for p53. Nat Genet. 1992 Apr;1(1):45–49. [PubMed]
  • Evans MJ, Kaufman MH. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981 Jul 9;292(5819):154–156. [PubMed]
  • Feinstein E, Gale RP, Reed J, Canaani E. Expression of the normal p53 gene induces differentiation of K562 cells. Oncogene. 1992 Sep;7(9):1853–1857. [PubMed]
  • Frebourg T, Barbier N, Kassel J, Ng YS, Romero P, Friend SH. A functional screen for germ line p53 mutations based on transcriptional activation. Cancer Res. 1992 Dec 15;52(24):6976–6978. [PubMed]
  • Friedlander P, Haupt Y, Prives C, Oren M. A mutant p53 that discriminates between p53-responsive genes cannot induce apoptosis. Mol Cell Biol. 1996 Sep;16(9):4961–4971. [PMC free article] [PubMed]
  • Funk WD, Pak DT, Karas RH, Wright WE, Shay JW. A transcriptionally active DNA-binding site for human p53 protein complexes. Mol Cell Biol. 1992 Jun;12(6):2866–2871. [PMC free article] [PubMed]
  • Gannon JV, Greaves R, Iggo R, Lane DP. Activating mutations in p53 produce a common conformational effect. A monoclonal antibody specific for the mutant form. EMBO J. 1990 May;9(5):1595–1602. [PMC free article] [PubMed]
  • Ginsberg D, Mechta F, Yaniv M, Oren M. Wild-type p53 can down-modulate the activity of various promoters. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):9979–9983. [PMC free article] [PubMed]
  • Gottlieb E, Haffner R, von Rüden T, Wagner EF, Oren M. Down-regulation of wild-type p53 activity interferes with apoptosis of IL-3-dependent hematopoietic cells following IL-3 withdrawal. EMBO J. 1994 Mar 15;13(6):1368–1374. [PMC free article] [PubMed]
  • Gottlieb E, Haffner R, King A, Asher G, Gruss P, Lonai P, Oren M. Transgenic mouse model for studying the transcriptional activity of the p53 protein: age- and tissue-dependent changes in radiation-induced activation during embryogenesis. EMBO J. 1997 Mar 17;16(6):1381–1390. [PMC free article] [PubMed]
  • Gottlieb TM, Oren M. p53 in growth control and neoplasia. Biochim Biophys Acta. 1996 Jun 7;1287(2-3):77–102. [PubMed]
  • Harris LC, Remack JS, Houghton PJ, Brent TP. Wild-type p53 suppresses transcription of the human O6-methylguanine-DNA methyltransferase gene. Cancer Res. 1996 May 1;56(9):2029–2032. [PubMed]
  • Harvey M, Sands AT, Weiss RS, Hegi ME, Wiseman RW, Pantazis P, Giovanella BC, Tainsky MA, Bradley A, Donehower LA. In vitro growth characteristics of embryo fibroblasts isolated from p53-deficient mice. Oncogene. 1993 Sep;8(9):2457–2467. [PubMed]
  • Haupt Y, Rowan S, Shaulian E, Vousden KH, Oren M. Induction of apoptosis in HeLa cells by trans-activation-deficient p53. Genes Dev. 1995 Sep 1;9(17):2170–2183. [PubMed]
  • Haupt Y, Maya R, Kazaz A, Oren M. Mdm2 promotes the rapid degradation of p53. Nature. 1997 May 15;387(6630):296–299. [PubMed]
  • Hunter T. Braking the cycle. Cell. 1993 Dec 3;75(5):839–841. [PubMed]
  • Hupp TR, Meek DW, Midgley CA, Lane DP. Regulation of the specific DNA binding function of p53. Cell. 1992 Nov 27;71(5):875–886. [PubMed]
  • Hupp TR, Sparks A, Lane DP. Small peptides activate the latent sequence-specific DNA binding function of p53. Cell. 1995 Oct 20;83(2):237–245. [PubMed]
  • Jacks T, Remington L, Williams BO, Schmitt EM, Halachmi S, Bronson RT, Weinberg RA. Tumor spectrum analysis in p53-mutant mice. Curr Biol. 1994 Jan 1;4(1):1–7. [PubMed]
  • Jones SN, Roe AE, Donehower LA, Bradley A. Rescue of embryonic lethality in Mdm2-deficient mice by absence of p53. Nature. 1995 Nov 9;378(6553):206–208. [PubMed]
  • Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 1991 Dec 1;51(23 Pt 1):6304–6311. [PubMed]
  • Kastan MB, Radin AI, Kuerbitz SJ, Onyekwere O, Wolkow CA, Civin CI, Stone KD, Woo T, Ravindranath Y, Craig RW. Levels of p53 protein increase with maturation in human hematopoietic cells. Cancer Res. 1991 Aug 15;51(16):4279–4286. [PubMed]
  • Kemp CJ, Wheldon T, Balmain A. p53-deficient mice are extremely susceptible to radiation-induced tumorigenesis. Nat Genet. 1994 Sep;8(1):66–69. [PubMed]
  • Knippschild U, Oren M, Deppert W. Abrogation of wild-type p53 mediated growth-inhibition by nuclear exclusion. Oncogene. 1996 Apr 18;12(8):1755–1765. [PubMed]
  • Knudson AG. Stem cell regulation, tissue ontogeny, and oncogenic events. Semin Cancer Biol. 1992 Jun;3(3):99–106. [PubMed]
  • Ko LJ, Prives C. p53: puzzle and paradigm. Genes Dev. 1996 May 1;10(9):1054–1072. [PubMed]
  • Kubbutat MH, Jones SN, Vousden KH. Regulation of p53 stability by Mdm2. Nature. 1997 May 15;387(6630):299–303. [PubMed]
  • Lechner MS, Mack DH, Finicle AB, Crook T, Vousden KH, Laimins LA. Human papillomavirus E6 proteins bind p53 in vivo and abrogate p53-mediated repression of transcription. EMBO J. 1992 Aug;11(8):3045–3052. [PMC free article] [PubMed]
  • Louis JM, McFarland VW, May P, Mora PT. The phosphoprotein p53 is down-regulated post-transcriptionally during embryogenesis in vertebrates. Biochim Biophys Acta. 1988 Sep 7;950(3):395–402. [PubMed]
  • Lu X, Lane DP. Differential induction of transcriptionally active p53 following UV or ionizing radiation: defects in chromosome instability syndromes? Cell. 1993 Nov 19;75(4):765–778. [PubMed]
  • Lu X, Park SH, Thompson TC, Lane DP. Ras-induced hyperplasia occurs with mutation of p53, but activated ras and myc together can induce carcinoma without p53 mutation. Cell. 1992 Jul 10;70(1):153–161. [PubMed]
  • Ludwig RL, Bates S, Vousden KH. Differential activation of target cellular promoters by p53 mutants with impaired apoptotic function. Mol Cell Biol. 1996 Sep;16(9):4952–4960. [PMC free article] [PubMed]
  • Lutzker SG, Levine AJ. A functionally inactive p53 protein in teratocarcinoma cells is activated by either DNA damage or cellular differentiation. Nat Med. 1996 Jul;2(7):804–810. [PubMed]
  • MacCallum DE, Hupp TR, Midgley CA, Stuart D, Campbell SJ, Harper A, Walsh FS, Wright EG, Balmain A, Lane DP, et al. The p53 response to ionising radiation in adult and developing murine tissues. Oncogene. 1996 Dec 19;13(12):2575–2587. [PubMed]
  • Mack DH, Vartikar J, Pipas JM, Laimins LA. Specific repression of TATA-mediated but not initiator-mediated transcription by wild-type p53. Nature. 1993 May 20;363(6426):281–283. [PubMed]
  • MacQueen HA. Lethality of radioisotopes in early mouse embryos. J Embryol Exp Morphol. 1979 Aug;52:203–208. [PubMed]
  • Maltzman W, Czyzyk L. UV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells. Mol Cell Biol. 1984 Sep;4(9):1689–1694. [PMC free article] [PubMed]
  • Martinez J, Georgoff I, Martinez J, Levine AJ. Cellular localization and cell cycle regulation by a temperature-sensitive p53 protein. Genes Dev. 1991 Feb;5(2):151–159. [PubMed]
  • Michalovitz D, Halevy O, Oren M. Conditional inhibition of transformation and of cell proliferation by a temperature-sensitive mutant of p53. Cell. 1990 Aug 24;62(4):671–680. [PubMed]
  • Milner J. Different forms of p53 detected by monoclonal antibodies in non-dividing and dividing lymphocytes. Nature. 1984 Jul 12;310(5973):143–145. [PubMed]
  • Milner J. Flexibility: the key to p53 function? Trends Biochem Sci. 1995 Feb;20(2):49–51. [PubMed]
  • Milner J, Medcalf EA. Cotranslation of activated mutant p53 with wild type drives the wild-type p53 protein into the mutant conformation. Cell. 1991 May 31;65(5):765–774. [PubMed]
  • Milner J, Watson JV. Addition of fresh medium induces cell cycle and conformation changes in p53, a tumour suppressor protein. Oncogene. 1990 Nov;5(11):1683–1690. [PubMed]
  • Morgenbesser SD, Williams BO, Jacks T, DePinho RA. p53-dependent apoptosis produced by Rb-deficiency in the developing mouse lens. Nature. 1994 Sep 1;371(6492):72–74. [PubMed]
  • Nevins JR. E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. Science. 1992 Oct 16;258(5081):424–429. [PubMed]
  • Oren M, Maltzman W, Levine AJ. Post-translational regulation of the 54K cellular tumor antigen in normal and transformed cells. Mol Cell Biol. 1981 Feb;1(2):101–110. [PMC free article] [PubMed]
  • Oren M, Reich NC, Levine AJ. Regulation of the cellular p53 tumor antigen in teratocarcinoma cells and their differentiated progeny. Mol Cell Biol. 1982 Apr;2(4):443–449. [PMC free article] [PubMed]
  • Parker SB, Eichele G, Zhang P, Rawls A, Sands AT, Bradley A, Olson EN, Harper JW, Elledge SJ. p53-independent expression of p21Cip1 in muscle and other terminally differentiating cells. Science. 1995 Feb 17;267(5200):1024–1027. [PubMed]
  • Perry ME, Levine AJ. Tumor-suppressor p53 and the cell cycle. Curr Opin Genet Dev. 1993 Feb;3(1):50–54. [PubMed]
  • Pierce GB, Speers WC. Tumors as caricatures of the process of tissue renewal: prospects for therapy by directing differentiation. Cancer Res. 1988 Apr 15;48(8):1996–2004. [PubMed]
  • Pratt MA, Kralova J, McBurney MW. A dominant negative mutation of the alpha retinoic acid receptor gene in a retinoic acid-nonresponsive embryonal carcinoma cell. Mol Cell Biol. 1990 Dec;10(12):6445–6453. [PMC free article] [PubMed]
  • Purdie CA, Harrison DJ, Peter A, Dobbie L, White S, Howie SE, Salter DM, Bird CC, Wyllie AH, Hooper ML, et al. Tumour incidence, spectrum and ploidy in mice with a large deletion in the p53 gene. Oncogene. 1994 Feb;9(2):603–609. [PubMed]
  • Rivas CI, Wisniewski D, Strife A, Perez A, Lambek C, Bruno S, Darzynkiewicz Z, Clarkson B. Constitutive expression of p53 protein in enriched normal human marrow blast cell populations. Blood. 1992 Apr 15;79(8):1982–1986. [PubMed]
  • Robertson E, Bradley A, Kuehn M, Evans M. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature. 1986 Oct 2;323(6087):445–448. [PubMed]
  • Rogel A, Popliker M, Webb CG, Oren M. p53 cellular tumor antigen: analysis of mRNA levels in normal adult tissues, embryos, and tumors. Mol Cell Biol. 1985 Oct;5(10):2851–2855. [PMC free article] [PubMed]
  • Ronen D, Schwartz D, Teitz Y, Goldfinger N, Rotter V. Induction of HL-60 cells to undergo apoptosis is determined by high levels of wild-type p53 protein whereas differentiation of the cells is mediated by lower p53 levels. Cell Growth Differ. 1996 Jan;7(1):21–30. [PubMed]
  • Rutledge JC, Shourbaji AG, Hughes LA, Polifka JE, Cruz YP, Bishop JB, Generoso WM. Limb and lower-body duplications induced by retinoic acid in mice. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5436–5440. [PMC free article] [PubMed]
  • Sabbatini P, Lin J, Levine AJ, White E. Essential role for p53-mediated transcription in E1A-induced apoptosis. Genes Dev. 1995 Sep 1;9(17):2184–2192. [PubMed]
  • Sah VP, Attardi LD, Mulligan GJ, Williams BO, Bronson RT, Jacks T. A subset of p53-deficient embryos exhibit exencephaly. Nat Genet. 1995 Jun;10(2):175–180. [PubMed]
  • Savatier P, Huang S, Szekely L, Wiman KG, Samarut J. Contrasting patterns of retinoblastoma protein expression in mouse embryonic stem cells and embryonic fibroblasts. Oncogene. 1994 Mar;9(3):809–818. [PubMed]
  • Savatier P, Lapillonne H, van Grunsven LA, Rudkin BB, Samarut J. Withdrawal of differentiation inhibitory activity/leukemia inhibitory factor up-regulates D-type cyclins and cyclin-dependent kinase inhibitors in mouse embryonic stem cells. Oncogene. 1996 Jan 18;12(2):309–322. [PubMed]
  • Schreiber M, Baumann B, Cotten M, Angel P, Wagner EF. Fos is an essential component of the mammalian UV response. EMBO J. 1995 Nov 1;14(21):5338–5349. [PMC free article] [PubMed]
  • Shaulsky G, Goldfinger N, Peled A, Rotter V. Involvement of wild-type p53 in pre-B-cell differentiation in vitro. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8982–8986. [PMC free article] [PubMed]
  • Sibilia M, Wagner EF. Strain-dependent epithelial defects in mice lacking the EGF receptor. Science. 1995 Jul 14;269(5221):234–238. [PubMed]
  • Soddu S, Blandino G, Citro G, Scardigli R, Piaggio G, Ferber A, Calabretta B, Sacchi A. Wild-type p53 gene expression induces granulocytic differentiation of HL-60 cells. Blood. 1994 Apr 15;83(8):2230–2237. [PubMed]
  • Soddu S, Blandino G, Scardigli R, Coen S, Marchetti A, Rizzo MG, Bossi G, Cimino L, Crescenzi M, Sacchi A. Interference with p53 protein inhibits hematopoietic and muscle differentiation. J Cell Biol. 1996 Jul;134(1):193–204. [PMC free article] [PubMed]
  • Symonds H, Krall L, Remington L, Saenz-Robles M, Lowe S, Jacks T, Van Dyke T. p53-dependent apoptosis suppresses tumor growth and progression in vivo. Cell. 1994 Aug 26;78(4):703–711. [PubMed]
  • Ueda H, Ullrich SJ, Gangemi JD, Kappel CA, Ngo L, Feitelson MA, Jay G. Functional inactivation but not structural mutation of p53 causes liver cancer. Nat Genet. 1995 Jan;9(1):41–47. [PubMed]
  • Vogelstein B, Kinzler KW. p53 function and dysfunction. Cell. 1992 Aug 21;70(4):523–526. [PubMed]
  • Wang QM, Jones JB, Studzinski GP. Cyclin-dependent kinase inhibitor p27 as a mediator of the G1-S phase block induced by 1,25-dihydroxyvitamin D3 in HL60 cells. Cancer Res. 1996 Jan 15;56(2):264–267. [PubMed]
  • Wang ZQ, Grigoriadis AE, Möhle-Steinlein U, Wagner EF. A novel target cell for c-fos-induced oncogenesis: development of chondrogenic tumours in embryonic stem cell chimeras. EMBO J. 1991 Sep;10(9):2437–2450. [PMC free article] [PubMed]
  • Weinberg WC, Azzoli CG, Chapman K, Levine AJ, Yuspa SH. p53-mediated transcriptional activity increases in differentiating epidermal keratinocytes in association with decreased p53 protein. Oncogene. 1995 Jun 15;10(12):2271–2279. [PubMed]
  • Wiman KG. The retinoblastoma gene: role in cell cycle control and cell differentiation. FASEB J. 1993 Jul;7(10):841–845. [PubMed]

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