Noncanonical DNA Motifs as Transactivation Targets by Wild Type and Mutant p53
Open accessAbstract
Sequence-specific binding by the human p53 master regulator is critical to its tumor suppressor activity in response to environmental stresses. p53 binds as a tetramer to two decameric half-sites separated by 0-13 nucleotides (nt), originally defined by the consensus RRRCWWGYYY (n = 0-13) RRRCWWGYYY. To better understand the role of sequence, organization, and level of p53 on transactivation at target response elements (REs) by wild type (WT) and mutant p53, we deconstructed the functional p53 canonical consensus sequence using budding yeast and human cell systems. Contrary to early reports on binding in vitro, small increases in distance between decamer half-sites greatly reduces p53 transactivation, as demonstrated for the natural TIGER RE. This was confirmed with human cell extracts using a newly developed, semi-in vitro microsphere binding assay. These results contrast with the synergistic increase in transactivation from a pair of weak, full-site REs in the MDM2 promoter that are separated by an evolutionary conserved 17 bp spacer. Surprisingly, there can be substantial transactivation at noncanonical (1/2)-(a single decamer) and (3/4)-sites, some of which were originally classified as biologically relevant canonical consensus sequences including PIDD and Apaf-1. p53 family members p63 and p73 yielded similar results. Efficient transactivation from noncanonical elements requires tetrameric p53, and the presence of the carboxy terminal, non-specific DNA binding domain enhanced transactivation from noncanonical sequences. Our findings demonstrate that RE sequence, organization, and level of p53 can strongly impact p53-mediated transactivation, thereby changing the view of what constitutes a functional p53 target. Importantly, inclusion of (1/2)- and (3/4)-site REs greatly expands the p53 master regulatory network.
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Results from multiple studies suggest that p53 can bind and transactivate target genes through noncanonical binding sites, particularly through half-sites.
“In addition, results from multiple studies suggest that p53 can bind and transactivate target genes through noncanonical binding sites, particularly through half-sites.22, 54, 55, 56”
From Census and evaluation of p53 target genes (2017) · cites this paper as [54]
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Self-citationIn vitro structure-function analysis of the β-strand 326-333 of human p53 1 1Edited by A. Fersht
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Self-citationAnalysis of p53 quaternary structure in relation to sequence-specific DNA binding.
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Self-citationFunctional Analysis of the Human p53 Tumor Suppressor and its Mutants Using Yeast
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Blinded by the Light: The Growing Complexity of p53
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Self-citationCaspase-2-Mediated Cleavage of Mdm2 Creates a p53-Induced Positive Feedback Loop
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Self-citationAutophagy and cardiac diseases: Therapeutic potential of natural products
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Self-citationRedefining the p53 response element
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Self-citationp53 responsive elements in human retrotransposons
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Self-citationWhole-genome cartography of p53 response elements ranked on transactivation potential
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Self-citationConservation of DNA-binding specificity and oligomerisation properties within the p53 family
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Self-citationMore targets, more pathways and more clues for mutant p53
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Self-citationTo repress or not to repress: This is the guardian's question
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Structure of p73 DNA-binding domain tetramer modulates p73 transactivation
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Self-citationGain-of-function p53 mutants have widespread genomic locations partially overlapping with p63
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p53 Binding to Nucleosomal DNA Depends on the Rotational Positioning of DNA Response Element
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Self-citationTP53 Mutants in the Tower of Babel of Cancer Progression
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Self-citationDiverse p53/DNA binding modes expand the repertoire of p53 response elements
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Self-citationTwo p53 tetramers bind one consensus DNA response element
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Low-level p53 expression changes transactivation rules and reveals superactivating sequences
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Self-citationNew Therapeutic Strategies for Cancer and Neurodegeneration Emerging from Yeast Cell-based Systems
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Self-citationSingle-Molecule Studies of Unlabeled Full-Length p53 Protein Binding to DNA
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Making Connections: p53 and the Cathepsin Proteases as Co-Regulators of Cancer and Apoptosis
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Self-citationMolecular Basis for Modulation of the p53 Target Selectivity by KLF4
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Sequence Properties of an Intramolecular Interaction that Inhibits p53 DNA Binding
Gregory, E., & Daughdrill, G. W. (2022) · Biomolecules · doi:10.3390/biom12111558 · Cited by 8 · see citing works
Regression based predictor for p53 transactivation
Gowrisankar, S., & Jegga, A. G. (2009) · BMC Bioinformatics · doi:10.1186/1471-2105-10-215 · Cited by 7 · see citing works
MOCCS: Clarifying DNA-binding motif ambiguity using ChIP-Seq data
Ozaki, H., & Iwasaki, W. (2016) · Computational Biology and Chemistry · doi:10.1016/j.compbiolchem.2016.01.014 · Cited by 6 · see citing works
Farooqi, K. M., Ghazvini, M., Pride, L. D., Mazzella, L., White, D. E., Pramanik, A., Bargonetti, J., & Moore, C. W. (2020) · Biomolecules · doi:10.3390/biom10030417 · Cited by 6 · see citing works
Menéndez, D., Resnick, M. A., & Haran, T. E. (2012) · Cell Cycle · doi:10.4161/cc.22467 · Cited by 6 · see citing works
Self-citationCai, B., Chao, C., Lin, H.-C., Huang, H., Kannagi, R., & Chen, J.-Y. (2016) · The Journal of Biochemistry · doi:10.1093/jb/mvw005 · Cited by 5 · see citing works
Quantitative Analysis of NF-κB Transactivation Specificity Using a Yeast-Based Functional Assay
Sharma, V., Jordan, J. J., Ciribilli, Y., Resnick, M. A., Bisio, A., & Inga, A. (2015) · PLoS ONE · doi:10.1371/journal.pone.0130170 · Cited by 5 · see citing works
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Involvement of p53 in the S-phase Checkpoint during Nucleotide Deficiencies
Heyer, C. L. (2011) · VCU Scholars Compass (Virginia Commonwealth University) · doi:10.25772/4zf5-2439
Modulation of p53's transcriptional function by small molecules
Nikulenkov, F. (2011) · Figshare
Predicting Functional Impact of Coding and Non-Coding Single Nucleotide Polymorphisms
Gowrisankar, S. (2008) · OhioLink ETD Center (Ohio Library and Information Network)
OPINION Interactions between the tumor suppressor p53 and immune responses
Menéndez, D., Shatz, M., & Resnick, M. A. (2013)
Self-citationJordan, J. J., Inga, A., Conway, K., Edmiston, S. N., Carey, L., Wu, L., & Resnick, M. A. (2023) · doi:10.1158/1541-7786.c.6542128
Self-citationJordan, J. J., Inga, A., Conway, K., Edmiston, S. N., Carey, L., Wu, L., & Resnick, M. A. (2023) · doi:10.1158/1541-7786.c.6542128.v1
Self-citationOptimisation of TP53 reporters by systematic dissection of synthetic TP53 response elementsPreprint
Trauernicht, M., Rastogi, C., Manzo, S. G., Bussemaker, H. J., & van Steensel, B. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.07.24.549988
Molecular characterization of FOX factors and Wnt signalling interplay in human cancers
Pizzolato, G. (2023) · Linköping University medical dissertations · doi:10.3384/9789180753364
Kanwar, N., Kumar, A. H. S., Gupta, N., Gagneja, G., & Kanwal, A. (2025) · Elsevier eBooks · doi:10.1016/b978-0-443-33343-9.00008-9
- Authors
- Jordan, J. J., Menendez, D., Inga, A., Nourredine, M., Bell, D., & Resnick, M. A.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- National Institutes of Health (US)
- Corresponding author
- Resnick, Michael A.
- Venue
- PLoS Genetics
- Publisher
- Public Library of Science (PLoS)
- Published
- 2008-06-27
- Volume
- 4
- Issue
- 6
- Pages
- e1000104
- ISSN
- 1553-7390 (ISSN-L)
- 1553-7404
- PMC
- PMC2518093
- OpenAlex
- W2049945082
- Cited by
- 103
- References
- 113
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 6 · 2022: 3 · 2021: 3 · 2020: 4 · 2019: 3 · 2018: 3 · 2017: 3 · 2016: 7 · 2015: 6 · 2014: 4 · 2013: 10 · 2012: 13
- FWCI
- 6.16
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS Genetics (publishedVersion, public-domain)
- PubMed Central (publishedVersion, public-domain)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Funders
- U.S. Department of Defense
- Associazione Italiana per la Ricerca sul Cancro
- National Institute of Environmental Health Sciences
- Topics
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- p53, transcriptional activation, response elements, p63, p73, p53 tetramer, tumor suppressor regulation
- MeSH terms
- Base Sequence, Binding Sites, Biological Assay, Diploidy, DNA, Humans, Molecular Sequence Data, Promoter Regions, Genetic, Saccharomyces cerevisiae, Transfection, Transcriptional Activation, Tumor Suppressor Protein p53, Consensus Sequence, Point Mutation, Protein Structure, Tertiary, Genes, Reporter, Dimerization, Response Elements, DNA, Intergenic, Cell Line, Tumor, Luciferases, Renilla, Proto-Oncogene Proteins c-mdm2
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