Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53
Open accessAbstract
The p53 tumor suppressor protein accumulates to very high concentrations in normal human fibroblasts infected by adenovirus type 5 mutants that cannot direct assembly of the viral E1B 55-kDa protein-containing E3 ubiquitin ligase that targets p53 for degradation. Despite high concentrations of nuclear p53, the p53 transcriptional program is not induced in these infected cells. We exploited this system to examine select post-translational modifications (PTMs) present on a transcriptionally inert population of endogenous human p53, as well as on p53 activated in response to etoposide treatment of normal human fibroblasts. These forms of p53 were purified from whole cell lysates by means of immunoaffinity chromatography and SDS-PAGE, and peptides derived from them were subjected to nano-ultra-high-performance LC-MS and MS/MS analyses on a high-resolution accurate-mass MS platform (data available via ProteomeXchange, PXD000464). We identified an unexpectedly large number of PTMs, comprising phosphorylation of Ser and Thr residues, methylation of Arg residues, and acetylation, ubiquitinylation, and methylation of Lys residues-for example, some 150 previously undescribed modifications of p53 isolated from infected cells. These modifications were distributed across all functional domains of both forms of the endogenous human p53 protein, as well as those of an orthologous population of p53 isolated from COS-1 cells. Despite the differences in activity, including greater in vitro sequence-specific DNA binding activity exhibited by p53 isolated from etoposide-treated cells, few differences were observed in the location, nature, or relative frequencies of PTMs on the two populations of human p53. Indeed, the wealth of PTMs that we have identified is consistent with a far greater degree of complex, combinatorial regulation of p53 by PTM than previously anticipated.
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PTMs of transcription factors represent highly tractable targets for modulation of TF activity.
“These modifications can alter the DNA binding activity and cofactor interactions of TFs, representing highly tractable targets for modulation of TF activity.643,644”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [643]
A 2014 proteomics study identified 222 PTMs of 99 residues on p53, a tumor suppressor protein.
“In 2014, a proteomics study identified 222 PTMs of 99 residues on p53, a tumor suppressor protein.”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [643]
PTMs can alter the DNA binding activity and cofactor interactions of transcription factors.
“These modifications can alter the DNA binding activity and cofactor interactions of TFs, representing highly tractable targets for modulation of TF activity.643,644”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [643]
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Self-citationCharacterization of NEK10 Tyrosine Kinase Activity in the Cellular Response to DNA Damage
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- Authors
- DeHart, C. J., Chahal, J. S., Flint, S. J., & Perlman, D. H.
- Affiliations
- Princeton University (US)
- Venue
- Molecular & Cellular Proteomics
- Publisher
- Elsevier BV
- Published
- 2014-01
- Volume
- 13
- Issue
- 1
- Pages
- 1-17
- ISSN
- 1535-9476 (ISSN-L)
- PMC
- PMC3879606
- OpenAlex
- W2122510296
- Cited by
- 70
- References
- 149
- Citations by year
- 2026: 5 · 2025: 1 · 2024: 7 · 2023: 5 · 2022: 3 · 2021: 5 · 2020: 3 · 2019: 8 · 2018: 8 · 2017: 5 · 2016: 4 · 2015: 9 · 2014: 7
- FWCI
- 3.57
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular & Cellular Proteomics (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- U.S. Public Health Service (R56A111091785)
- National Institute of Allergy and Infectious Diseases (R01AI1058172, R01AI058172, R56AI091785)
- Topics
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- p53, post-translational modification, p53 inactivation, p53 phosphorylation, lysine methylation, lysine acetylation, ubiquitination, arginine methylation, etoposide, E3 ubiquitin ligase, p53 transcriptional activation, p53 DNA binding, tumor suppressor regulation
- MeSH terms
- Acetylation, Animals, Chlorocebus aethiops, DNA-Binding Proteins, Fibroblasts, Humans, Methylation, Phosphorylation, Protein Processing, Post-Translational, Tumor Suppressor Protein p53, Adenovirus E1B Proteins, COS Cells, Ubiquitin-Protein Ligases, Proteolysis
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