Mouse models to investigate in situ cell fate decisions induced by p53.
Apoptosis
Cancer
Cell Cycle Arrest
Reporter Mice
p53/TRP53/TP53
Journal
The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664
Informations de publication
Date de publication:
19 Aug 2024
19 Aug 2024
Historique:
received:
19
06
2023
accepted:
12
07
2024
revised:
02
07
2024
medline:
20
8
2024
pubmed:
20
8
2024
entrez:
19
8
2024
Statut:
aheadofprint
Résumé
Investigating how transcription factors control complex cellular processes requires tools that enable responses to be visualised at the single-cell level and their cell fate to be followed over time. For example, the tumour suppressor p53 (also called TP53 in humans and TRP53 in mice) can initiate diverse cellular responses by transcriptional activation of its target genes: Puma to induce apoptotic cell death and p21 to induce cell cycle arrest/cell senescence. However, it is not known how these processes are regulated and initiated in different cell types. Also, the context-dependent interaction partners and binding loci of p53 remain largely elusive. To be able to examine these questions, we here developed knock-in mice expressing triple-FLAG-tagged p53 to facilitate p53 pull-down and two p53 response reporter mice, knocking tdTomato and GFP into the Puma/Bbc3 and p21 gene loci, respectively. By crossing these reporter mice into a p53-deficient background, we show that the new reporters reliably inform on p53-dependent and p53-independent initiation of both apoptotic or cell cycle arrest/senescence programs, respectively, in vitro and in vivo.
Identifiants
pubmed: 39160273
doi: 10.1038/s44318-024-00189-z
pii: 10.1038/s44318-024-00189-z
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 1154970
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 1113133
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 1116937
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 1143105
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 2002618
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 2001201
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 2011139
Organisme : DHAC | National Health and Medical Research Council (NHMRC)
ID : 2010275
Organisme : Victorian Cancer Agency (VCA)
ID : 17028
Informations de copyright
© 2024. The Author(s).
Références
Amemiya HM, Kundaje A, Boyle AP (2019) The ENCODE blacklist: identification of problematic regions of the genome. Sci Rep 9:9354
pubmed: 31249361
pmcid: 6597582
doi: 10.1038/s41598-019-45839-z
Boutelle AM, Attardi LD (2021) p53 and tumor suppression: it takes a network. Trends Cell Biol 31:298–310
pubmed: 33518400
pmcid: 7954925
doi: 10.1016/j.tcb.2020.12.011
Brady CA, Jiang D, Mello SS, Johnson TM, Jarvis LA, Kozak MM, Kenzelmann Broz D, Basak S, Park EJ, McLaughlin ME et al (2011) Distinct p53 transcriptional programs dictate acute DNA-damage responses and tumor suppression. Cell 145:571–583
pubmed: 21565614
pmcid: 3259909
doi: 10.1016/j.cell.2011.03.035
Clarke AR, Purdie CA, Harrison DJ, Morris RG, Bird CC, Hooper ML, Wyllie AH (1993) Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 362:849–852
pubmed: 8479523
doi: 10.1038/362849a0
de Sousa Abreu R, Penalva LO, Marcotte EM, Vogel C (2009) Global signatures of protein and mRNA expression levels. Mol Biosyst 5:1512–1526
pubmed: 20023718
Delbridge ARD, Kueh AJ, Ke F, Zamudio NM, El-Saafin F, Jansz N, Wang GY, Iminitoff M, Beck T, Haupt S et al (2019) Loss of p53 causes stochastic aberrant X-chromosome inactivation and female-specific neural tube defects. Cell Rep 27:442–454 e445
pubmed: 30970248
doi: 10.1016/j.celrep.2019.03.048
Deng C, Zhang P, Harper JW, Elledge SJ, Leder P (1995) Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82:675–684
pubmed: 7664346
doi: 10.1016/0092-8674(95)90039-X
Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery Jr. CA, Butel JS, Bradley A (1992) Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature 356:215–221
pubmed: 1552940
doi: 10.1038/356215a0
el-Deiry WS, Kern SE, Pietenpol JA, Kinzler KW, Vogelstein B (1992) Definition of a consensus binding site for p53. Nat Genet 1:45–49
pubmed: 1301998
doi: 10.1038/ng0492-45
Fischer M (2017) Census and evaluation of p53 target genes. Oncogene 36:3943–3956
pubmed: 28288132
pmcid: 5511239
doi: 10.1038/onc.2016.502
Fischer M, Grossmann P, Padi M, DeCaprio JA (2016) Integration of TP53, DREAM, MMB-FOXM1 and RB-E2F target gene analyses identifies cell cycle gene regulatory networks. Nucleic Acids Res 44:6070–6086
pubmed: 27280975
pmcid: 4994865
doi: 10.1093/nar/gkw523
Funk WD, Pak DT, Karas RH, Wright WE, Shay JW (1992) A transcriptionally active DNA-binding site for human p53 protein complexes. Mol Cell Biol 12:2866–2871
pubmed: 1588974
pmcid: 364481
Goh AM, Lim CY, Chiam PC, Li L, Mann MB, Mann KM, Menendez S, Lane DP (2012) Using targeted transgenic reporter mice to study promoter-specific p53 transcriptional activity. Proc Natl Acad Sci USA 109:1685–1690
pubmed: 22307631
pmcid: 3277193
doi: 10.1073/pnas.1114173109
Hafner A, Kublo L, Tsabar M, Lahav G, Stewart-Ornstein J (2020) Identification of universal and cell-type specific p53 DNA binding. BMC Mol Cell Biol 21:5
pubmed: 32070277
pmcid: 7027055
doi: 10.1186/s12860-020-00251-8
Hahne F, Ivanek R (2016) Visualizing genomic data using Gviz and Bioconductor. Methods Mol Biol 1418:335–351
pubmed: 27008022
doi: 10.1007/978-1-4939-3578-9_16
Han J, Flemington C, Houghton AB, Gu Z, Zambetti GP, Lutz RJ, Zhu L, Chittenden T (2001) Expression of bbc3, a pro-apoptotic BH3-only gene, is regulated by diverse cell death and survival signals. Proc Natl Acad Sci USA 98:11318–11323
pubmed: 11572983
pmcid: 58727
doi: 10.1073/pnas.201208798
Hawkins ED, Duarte D, Akinduro O, Khorshed RA, Passaro D, Nowicka M, Straszkowski L, Scott MK, Rothery S, Ruivo N et al (2016) T-cell acute leukaemia exhibits dynamic interactions with bone marrow microenvironments. Nature 538:518–522
pubmed: 27750279
pmcid: 5164929
doi: 10.1038/nature19801
Hemann MT, Zilfou JT, Zhao Z, Burgess DJ, Hannon GJ, Lowe SW (2004) Suppression of tumorigenesis by the p53 target PUMA. Proc Natl Acad Sci USA 101:9333–9338
pubmed: 15192153
pmcid: 438977
doi: 10.1073/pnas.0403286101
Jacks T, Remington L, Williams BO, Schmitt EM, Halachmi S, Bronson RT, Weinberg RA (1994) Tumor spectrum analysis in p53-mutant mice. Curr Biol 4:1–7
pubmed: 7922305
doi: 10.1016/S0960-9822(00)00002-6
Janic A, Valente LJ, Wakefield MJ, Di Stefano L, Milla L, Wilcox S, Yang H, Tai L, Vandenberg CJ, Kueh AJ et al (2018) DNA repair processes are critical mediators of p53-dependent tumor suppression. Nat Med 24:947–953
pubmed: 29892060
doi: 10.1038/s41591-018-0043-5
Jeffers JR, Parganas E, Lee Y, Yang C, Wang J, Brennan J, MacLean KH, Han J, Chittenden T, Ihle JN et al (2003) Puma is an essential mediator of p53-dependent and -independent apoptotic pathways. Cancer Cell 4:321–328
pubmed: 14585359
doi: 10.1016/S1535-6108(03)00244-7
Kastenhuber ER, Lowe SW (2017) Putting p53 in context. Cell 170:1062–1078
pubmed: 28886379
pmcid: 5743327
doi: 10.1016/j.cell.2017.08.028
Kenzelmann Broz D, Spano Mello S, Bieging KT, Jiang D, Dusek RL, Brady CA, Sidow A, Attardi LD (2013) Global genomic profiling reveals an extensive p53-regulated autophagy program contributing to key p53 responses. Genes Dev 27:1016–1031
pubmed: 23651856
pmcid: 3656320
doi: 10.1101/gad.212282.112
Levine AJ (2020) p53: 800 million years of evolution and 40 years of discovery. Nat Rev Cancer 20:471–480
pubmed: 32404993
doi: 10.1038/s41568-020-0262-1
Li Q, Mao F, Zhou B, Huang Y, Zou Z, denDekker AD, Xu J, Hou S, Liu J, Dou Y et al (2020) p53 integrates temporal WDR5 inputs during neuroectoderm and mesoderm differentiation of mouse embryonic stem cells. Cell Rep 30:465–480 e466
pubmed: 31940490
pmcid: 7024586
doi: 10.1016/j.celrep.2019.12.039
Li T, Kon N, Jiang L, Tan M, Ludwig T, Zhao Y, Baer R, Gu W (2012) Tumor suppression in the absence of p53-mediated cell-cycle arrest, apoptosis, and senescence. Cell 149:1269–1283
pubmed: 22682249
pmcid: 3688046
doi: 10.1016/j.cell.2012.04.026
Liao Y, Smyth GK, Shi W (2019) The R package Rsubread is easier, faster, cheaper and better for alignment and quantification of RNA sequencing reads. Nucleic Acids Res 47:e47
pubmed: 30783653
pmcid: 6486549
doi: 10.1093/nar/gkz114
Lieschke E, Wang Z, Chang C, Weeden CE, Kelly GL, Strasser A (2022) Flow cytometric single cell-based assay to simultaneously detect cell death, cell cycling, DNA content and cell senescence. Cell Death Differ 29:1004–1012
pubmed: 35264779
pmcid: 9091206
doi: 10.1038/s41418-022-00964-7
Lotem J, Sachs L (1993) Hematopoietic cells from mice deficient in wild-type p53 are more resistant to induction of apoptosis by some agents. Blood 82:1092–1096
pubmed: 8353276
doi: 10.1182/blood.V82.4.1092.1092
Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T (1993) p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362:847–849
pubmed: 8479522
doi: 10.1038/362847a0
McMahon M, Frangova TG, Henderson CJ, Wolf CR (2016) Olaparib, monotherapy or with ionizing radiation, exacerbates DNA damage in normal tissues: insights from a new p21 reporter mouse. Mol Cancer Res 14:1195–1203
pubmed: 27604276
pmcid: 5136472
doi: 10.1158/1541-7786.MCR-16-0108
Michalak EM, Jansen ES, Happo L, Cragg MS, Tai L, Smyth GK, Strasser A, Adams JM, Scott CL (2009) Puma and to a lesser extent Noxa are suppressors of Myc-induced lymphomagenesis. Cell Death Differ 16:684–696
pubmed: 19148184
doi: 10.1038/cdd.2008.195
Michalak EM, Villunger A, Adams JM, Strasser A (2008) In several cell types tumour suppressor p53 induces apoptosis largely via Puma but Noxa can contribute. Cell Death Differ 15:1019–1029
pubmed: 18259198
doi: 10.1038/cdd.2008.16
Moyer SM, Wasylishen AR, Qi Y, Fowlkes N, Su X, Lozano G (2020) p53 drives a transcriptional program that elicits a non-cell-autonomous response and alters cell state in vivo. Proc Natl Acad Sci USA 117:23663–23673
pubmed: 32900967
pmcid: 7519296
doi: 10.1073/pnas.2008474117
Nakano K, Vousden KH (2001) PUMA, a novel proapoptotic gene, is induced by p53. Mol Cell 7:683–694
pubmed: 11463392
doi: 10.1016/S1097-2765(01)00214-3
Ramirez F, Ryan DP, Gruning B, Bhardwaj V, Kilpert F, Richter AS, Heyne S, Dundar F, Manke T (2016) deepTools2: a next generation web server for deep-sequencing data analysis. Nucleic Acids Res 44:W160–165
pubmed: 27079975
pmcid: 4987876
doi: 10.1093/nar/gkw257
Resnick-Silverman L, Zhou R, Campbell MJ, Leibling I, Parsons R, Manfredi JJ (2023) In vivo RNA-seq and ChIP-seq analyses show an obligatory role for the C terminus of p53 in conferring tissue-specific radiation sensitivity. Cell Rep 42:112216
pubmed: 36924496
pmcid: 10200257
doi: 10.1016/j.celrep.2023.112216
Riley T, Sontag E, Chen P, Levine A (2008) Transcriptional control of human p53-regulated genes. Nat Rev Mol Cell Biol 9:402–412
pubmed: 18431400
doi: 10.1038/nrm2395
Sanchez Y, Segura V, Marin-Bejar O, Athie A, Marchese FP, Gonzalez J, Bujanda L, Guo S, Matheu A, Huarte M (2014) Genome-wide analysis of the human p53 transcriptional network unveils a lncRNA tumour suppressor signature. Nat Commun 5:5812
pubmed: 25524025
doi: 10.1038/ncomms6812
Šidák ZK (1967) Rectangular confidence regions for the means of multivariate normal distributions. J Am Stat Assoc 62:626–633
Strasser A, Harris AW, Cory S (1991) bcl-2 transgene inhibits T cell death and perturbs thymic self-censorship. Cell 67:889–899
pubmed: 1959134
doi: 10.1016/0092-8674(91)90362-3
Strasser A, Harris AW, Jacks T, Cory S (1994) DNA damage can induce apoptosis in proliferating lymphoid cells via p53-independent mechanisms inhibitable by Bcl-2. Cell 79:329–339
pubmed: 7954799
doi: 10.1016/0092-8674(94)90201-1
Thomas AF, Kelly GL, Strasser A (2022) Of the many cellular responses activated by TP53, which ones are critical for tumour suppression? Cell Death Differ 29:961–971
pubmed: 35396345
pmcid: 9090748
doi: 10.1038/s41418-022-00996-z
Tinkum KL, Marpegan L, White LS, Sun J, Herzog ED, Piwnica-Worms D, Piwnica-Worms H (2011) Bioluminescence imaging captures the expression and dynamics of endogenous p21 promoter activity in living mice and intact cells. Mol Cell Biol 31:3759–3772
pubmed: 21791610
pmcid: 3165732
doi: 10.1128/MCB.05243-11
Torgovnick A, Heger JM, Liaki V, Isensee J, Schmitt A, Knittel G, Riabinska A, Beleggia F, Laurien L, Leeser U et al (2018) The Cdkn1a(SUPER) mouse as a tool to study p53-mediated tumor suppression. Cell Rep 25:1027–1039 e1026
pubmed: 30355482
doi: 10.1016/j.celrep.2018.09.079
Valente LJ, Aubrey BJ, Herold MJ, Kelly GL, Happo L, Scott CL, Newbold A, Johnstone RW, Huang DC, Vassilev LT et al (2016) Therapeutic response to non-genotoxic activation of p53 by Nutlin3a is driven by PUMA-mediated apoptosis in lymphoma cells. Cell Rep 14:1858–1866
pubmed: 26904937
doi: 10.1016/j.celrep.2016.01.059
Valente LJ, Gray DH, Michalak EM, Pinon-Hofbauer J, Egle A, Scott CL, Janic A, Strasser A (2013) p53 efficiently suppresses tumor development in the complete absence of its cell-cycle inhibitory and proapoptotic effectors p21, Puma, and Noxa. Cell Rep 3:1339–1345
pubmed: 23665218
doi: 10.1016/j.celrep.2013.04.012
Vasey DB, Wolf CR, MacArtney T, Brown K, Whitelaw CB (2008) p21-LacZ reporter mice reflect p53-dependent toxic insult. Toxicol Appl Pharmacol 227:440–450
pubmed: 18215733
doi: 10.1016/j.taap.2007.11.029
Vassilev LT, Vu BT, Graves B, Carvajal D, Podlaski F, Filipovic Z, Kong N, Kammlott U, Lukacs C, Klein C et al (2004) In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science 303:844–848
pubmed: 14704432
doi: 10.1126/science.1092472
Villunger A, Michalak EM, Coultas L, Mullauer F, Bock G, Ausserlechner MJ, Adams JM, Strasser A (2003) p53- and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa. Science 302:1036–1038
pubmed: 14500851
doi: 10.1126/science.1090072
Vogel C, Marcotte EM (2012) Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet 13:227–232
pubmed: 22411467
pmcid: 3654667
doi: 10.1038/nrg3185
Vousden KH, Lane DP (2007) p53 in health and disease. Nat Rev Mol Cell Biol 8:275–283
pubmed: 17380161
doi: 10.1038/nrm2147
Yu J, Zhang L, Hwang PM, Kinzler KW, Vogelstein B (2001) PUMA induces the rapid apoptosis of colorectal cancer cells. Mol Cell 7:673–682
pubmed: 11463391
doi: 10.1016/S1097-2765(01)00213-1
Zhang Y, Liu T, Meyer CA, Eeckhoute J, Johnson DS, Bernstein BE, Nusbaum C, Myers RM, Brown M, Li W et al (2008) Model-based analysis of ChIP-Seq (MACS). Genome Biol 9:R137
pubmed: 18798982
pmcid: 2592715
doi: 10.1186/gb-2008-9-9-r137