Identification and Characterization of a Novel p300-mediated p53 Acetylation Site, Lysine 305
Open accessAbstract
Post-translational modifications serve as important regulatory elements in modulating the transcriptional activity of the tumor suppressor protein p53. We have previously reported a tandem mass spectrometry-based method (viz. selected ion tracing analysis) that can be applied to the identification of phosphopeptides as well as exact mapping of the phosphorylated residues within. In this study, we describe the application of the same strategy for the identification of p300 acetyltransferase-mediated acetylation sites on p53. Consistent with the previous finding, lysines 370, 372, 373, 381, and 382 were detected by this modified selected ion tracing method as the target sites of p300 in vitro. Moreover, two novel acetylation sites, Lys-292 and Lys-305, were also found. Immunoblotting using anti-acetyl-Lys-305 antibody confirmed this discovery and demonstrated that Lys-305 could be acetylated by p300 both in vitro and in vivo. We also show that an alanine or glutamine substitution at Lys-305 (K305A or K305Q) suppressed the transcriptional activity of p53, whereas an arginine mutation (K305R) increased the transcriptional activity. Thus, p300 may further regulate the transcriptional activity of p53 through a novel acetylation site, Lys-305.
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Arginine is known to mimic a deacetylation status.
“In contrast to alanine, arginine is known to mimic a deacetylation status (16, 21, 25).”
From Identification of a Key Lysine Residue in Heat Shock Protein 90 Required for Azole and Echinocandin Resistance in Aspergillus fumigatus (2014) · cites this paper as [25]
Mutation of lysine to alanine is expected to mimic acetylation.
“Mutation of lysine to alanine is expected to mimic acetylation (16, 21, 25).”
From Identification of a Key Lysine Residue in Heat Shock Protein 90 Required for Azole and Echinocandin Resistance in Aspergillus fumigatus (2014) · cites this paper as [25]
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- Authors
- Wang, Y.-H., Tsay, Y.-G., Tan, B. C.-M., Lo, W.-Y., & Lee, S.-C.
- Affiliations
- National Taiwan University (TW)
- Corresponding author
- Lee, Sheng-Chung
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2003-07
- Volume
- 278
- Issue
- 28
- Pages
- 25568-25576
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2081774189
- Cited by
- 92
- References
- 37
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 2 · 2022: 3 · 2021: 4 · 2020: 2 · 2019: 4 · 2018: 4 · 2017: 4 · 2016: 4 · 2015: 6 · 2014: 4 · 2013: 3 · 2012: 4
- FWCI
- 2.69
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Science Council
- Academia Sinica
- 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 › Genomics and Chromatin Dynamics
- Keywords
- p53, P300, lysine acetylation, acetylation sites, post-translational modification, transcriptional activity, tumor suppressor regulation, site-directed mutagenesis
- MeSH terms
- p300-CBP-Associated Factor, Acetyltransferases, Alanine, Amino Acid Sequence, Butyrates, DNA, Glutamine, Humans, Ions, Lysine, Molecular Sequence Data, Peptides, Plasmids, Protein Binding, Mass Spectrometry, Time Factors, Transcription Factors, Transcription, Genetic, Transfection, Tumor Cells, Cultured, Ultraviolet Rays, Immunoblotting, Blotting, Western, Tumor Suppressor Protein p53, Mutagenesis, Site-Directed, Genes, Reporter, Cell Cycle Proteins, p300-CBP Transcription Factors, Histone Acetyltransferases, Isobutyrates
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