p53 Poster
p53 Poster
p53 Poster
1989 onwards
p53 has a pivotal role in human tumorigenesis
The turning point in p53 research came in 1989. The remaining TP53 allele sequenced from a tumour that had lost a portion of the p arm of chromosome 17, where TP53 resides, was found to be mutated. The point mutation (shown in the chromatogram below) was not found in normal tissue samples taken from the patient. This, coupled with the frequent loss of 17p in tumours, indicated that TP53 was a tumour suppressor gene. It became clear that the wild-type TP53 gene used to demonstrate that p53 was oncogenic in cell transformation experiments was, in fact, mutated. That p53 is a bona fide tumour suppressor was confirmed in 1990 by the finding that patients with LiFraumeni syndrome which predisposes to diverse tumour types had inherited TP53 mutations, and further confirmed in 1992 by experiments showing that Trp53 (which encodes mouse p53) knockout mice are prone to tumours.
Wild-type TP53 G C C C T G T G C A G C T G T G G
1991 onwards
The biological effects of p53
The first clue into the outcome of activating p53 was revealed when it was shown that the expression of wild-type p53 arrests the cell cycle in a subset of cell lines. It was subsequently shown that the expression of wild-type p53 in other cells results in cell death rather than arrest. These findings were major stimulants to the developing field of apoptosis as they showed that the regulation of apoptosis and the regulation of cell proliferation were equally important to tumour suppression. In the early 1990s two seminal observations were made: that p53 has a transactivation domain, and that it can bind to specific DNA sequences. The crystal structure of the p53 core domain bound to DNA showed exactly which p53 residues make contact with DNA and revealed how different tumourassociated mutations abrogate this binding. It is now understood that wild-type p53 functions as a tetrameric transcription factor that regulates net cell growth, inhibiting the cell cycle in some circumstances and promoting apoptosis in others, through the activation or repression of key target genes. It has also been suggested that p53 promotes apoptosis in the cytoplasm through mechanisms that do not involve transcription. Both apoptosis and cell cycle arrest have since been shown to be important for limiting the propagation of mutations following many types of cellular stress, especially DNA damage. Moreover, p53regulated genes that function in other cellular processes, such as senescence, metabolism, autophagy, angiogenesis and DNA repair, have also been identified and these might have a role in its tumour suppressive activities (not shown in the figure).
Cell stress DNA tumour virus rNTP depletion DNA damage Spindle damage Oncogene activation SV40, HPV or adenovirus Hypoxia
The Timeline arrow represents a graph of the number of publications on p53 each year since 1979 (not to scale).
2009
1999
2000 onwards
Mutant TP53
ATM
ATR RB P
Subsequent studies have demonstrated that TP53 is more frequently mutated in human tumours than any other gene in the genome, and >25,000 TP53 mutations have been reported to date. Of these mutations, 75% occur as missense mutations that predominantly occur in the DBD. The figure below shows the distribution of tumourassociated missense mutations relative to the domains of p53 and highlights the six most common mutations. These mutations can affect the structure of p53 (distorting or unfolding the native conformation) and they can impair DNA binding. Experimental models have revealed that such tumour-derived mutations can have dominant-negative properties and, in some cases, can confer gain-of-oncogenic function properties in the absence of wild-type p53.
Number of mutations
1,800 1,200 600 0
PRIMA-1, MIRA-1, CP-31398, STIMA-1 HL198C Nutlin, NU8354, chlorofusin, RITA, MI-219, BDP 23 GEM240 Tenovin-6
CHK2 CHK1
*Gendicine is already being given to cancer patients in China, following approval by the Chinese Food and Drug Administration in 2004.
Much remains unknown about this multi-talented tumour suppressor. For example, which changes in the microenvironment favour the selection of cells with TP53 mutations? Does continuous or unrepairable DNA damage or the presence of reactive oxygen species, perhaps in association with alternating cycles of hypoxia and normoxia, influence the survival of cells with TP53 mutations? We also do not understand why the expression of wild-type p53 results in apoptosis in some cells and cell cycle arrest in others, or how the various p53 post-translational modifications might regulate this switch. Finally, and perhaps most importantly, we do not yet know how to use our knowledge of p53 for therapeutic purposes. Approaches to reactivate mutant p53 or remove inhibition of wild-type p53 are being developed and some show great promise (see the TABLE). However, the field is wide open to new, creative approaches that effectively translate p53 to the clinic. p53 research lives on.
MDM2 MDM4
DAXX USP7
Proteasome
1993 onwards
10 8 6 4 2
50
100
150
200
250
300
350
393
TA
High levels of mutant p53 were detected in human intestinal tumour cells (brown) but not normal cells by immunohistochemistry.
PR
DBD
Tet Reg
Mutations in green affect DNA contacts, those in red cause local distortions and those in purple cause global denaturation. The x-axis indicates amino acid number.
Cellular stress or DNA tumour virus infection triggers mediating proteins to activate p53 by phosphorylating it or inhibiting its ubiquitylation by MDM2. Following further post-translational modifications, the p53 tetramer regulates transcription by binding to p53 response elements and recruiting cofactors, and the protein products lead to cell cycle arrest or apoptosis.
TA S33 P T18 P
1979
About Roche
Headquartered in Basel, Switzerland, Roche is a leader in research-focused healthcare with combined strengths in pharmaceuticals and diagnostics. Roche is the worlds largest biotech company with truly differentiated medicines in oncology, virology, inflammation, metabolism and the central nervous system. Roche is also the world leader in in vitro diagnostics, tissue-based cancer diagnostics and a pioneer in diabetes management. Roches personalized healthcare strategy aims at providing medicines and diagnostic tools that enable tangible improvements in the health, quality of life and survival of patients. In 2008 Roche had over 80,000 employees worldwide and invested almost 9 billion Swiss francs in R&D. Genentech, United States, is a wholly owned member of the Roche Group. Roche has a majority stake in Chugai Pharmaceutical, Japan.
1989
Contact information
Bert Vogelstein, The Ludwig Center for Cancer Genetics and Therapeutics, Howard Hughes Medical Institute and Sidney Kimmel Cancer Center at the Johns Hopkins Medical Institutions, Baltimore, Maryland, USA. Carol Prives, Department of Biological Sciences, Columbia University, New York, NY 10027, USA. e-mails: bertvog@gmail.com; clp3@columbia.edu
Specific residues are modified as shown, with phosphorylation (P) in orange, acetylation (A) in green, ubiquitylation (Ub) in dark blue, neddylation (N) in pink, methylation (M) in blue and sumoylation (SU) in yellow.
It was known as early as 1981 that in normal, unstressed cells the level of p53 protein is low owing to its rapid turnover, whereas high levels of p53 are observed in tumours. In 1993 MDM2 was found to be a transcriptional target of p53 and was subsequently shown to be an E3 ubiquitin ligase that targets p53 for degradation, defining a negative feedback loop. Then, in 2001 MDM4, an MDM2 homologue that was first identified in 1996, was also shown to be important for the regulation of p53 activity, although its precise role remains unclear. In addition to ubiquitylation by MDM2, p53 is subject to many other post-translational modifications in response to stress, as shown in the figure. These modifications modulate the activity of p53 and its binding partners and are thought to determine the resulting p53-dependent cellular response.
% of missense mutations
Further information
IARC TP53 database: http://www-p53.iarc.fr IARC Mouse models with targeted p53 alterations: http://www-p53.iarc.fr/MouseModelView.asp Nature Reviews Cancer Focus on p53 30 years on: http://www.nature.com/nrc/focus/p53.html Pathway Interaction Database for p53: http://pid.nci.nih.gov/search/pathway_landing.shtm l?what=graphic&jpg=on&pathway_ id=p53regulationpathway
Abbreviations
ATM, ataxia telangiectasia mutated; ATR, ataxia telangiectasia and Rad3 related; BAX, BCL2-associated X protein; BBC3, BCL2 binding component 3 (also known as PUMA); BDP, benzodiazepine; CDC25C, cell division cycle 25 homolog C; CDK, cyclin-dependent kinase; CDKN1A, cyclin-dependent kinase inhibitor 1A; CHK, checkpoint kinase; DAXX, death-domain associated protein; DBD, DNA-binding domain; HAT, histone acetyltransferase; HDAC2, histone deacetylase 2; HPV, human papillomavirus; MIRA-1, mutant p53 reactivation and induction of rapid apoptosis; PMAIP1, phorbol-12-myristate-13-acetate-induced protein 1 (also known as NOXA); PR, proline-rich domain; PRIMA-1, p53 reactivation and induction of massive apoptosis; RB, retinoblastoma; RE, response element;
Reg, carboxy-terminal regulatory domain; RITA, reactivation of p53 and induction of tumour cell apoptosis; rNTP, ribonucleotide triphosphate; SFN, stratifin; SV40, simian virus 40; TA, transactivation domain; Tet, tetramerization domain; TNFSF10, tumour necrosis factor (ligand) superfamily, member 10 (also known as TRAIL); TP53I3, tumour protein p53 inducible protein 3; USP7, ubiquitin carboxyl-terminal hydrolase 7. We are grateful to Varda Rotter and Ran Brosh for donating the immunohistochemistry image. Poster design by Lara Crow, edited by Gemma K. Alderton, Katharine H. Wrighton and Nicola McCarthy, copyedited by Meera Swami. 2009 Nature Publishing Group.