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Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Modulation of p53 function by SET8-mediated methylation at lysine 382
Xiaobing Shi1, Ioulia Kachirskaia, Hiroshi Yamaguchi
1Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA.
Molecular Cell
|August 21, 2007
Summary
The lysine methyltransferase SETD8 regulates the tumor suppressor protein p53 by monomethylating it at lysine 382. This modification suppresses p53
Area of Science:
- Biochemistry
- Molecular Biology
- Epigenetics
Background:
- Histone methylation is a key epigenetic mechanism regulating gene expression.
- Lysine methylation is increasingly recognized on nonhistone proteins, impacting diverse cellular functions.
- The SET8/PR-Set7 enzyme is a known lysine methyltransferase with roles in chromatin regulation.
Purpose of the Study:
- To investigate the role of the lysine methyltransferase SET8 in regulating the tumor suppressor protein p53.
- To identify specific methylation sites on p53 and their functional consequences.
- To elucidate the impact of SET8-mediated p53 methylation on p53-dependent cellular processes.
Main Methods:
- Chromatin immunoprecipitation followed by sequencing (ChIP-seq).
- Western blotting using specific antibodies against methylated p53.
- Quantitative real-time PCR (qRT-PCR) to assess gene expression.
- Depletion studies using siRNA or shRNA to reduce SET8 levels.
- Analysis of p53-mediated apoptosis and cell cycle checkpoint activation.
Main Results:
- SET8 specifically monomethylates p53 at lysine 382 (p53K382me1).
- p53K382me1 robustly suppresses the transcriptional activation of highly responsive p53 target genes.
- Depletion of SET8 enhances p53's proapoptotic and checkpoint activation functions.
- SET8 expression is downregulated in response to DNA damage.
Conclusions:
- SET8 acts as a p53-modifying enzyme, regulating its function through methylation.
- p53K382me1 is a novel posttranslational modification that modulates p53 activity.
- This study reveals a new layer of epigenetic regulation controlling p53-mediated tumor suppression and DNA damage response.
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