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Regulation of E2F1 activity by acetylation.
M A Martínez-Balbás1, U M Bauer, S J Nielsen
1Wellcome/CRC Institute and Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
The EMBO Journal
|February 17, 2000
Summary
Acetylation modifies E2F1, enhancing its DNA-binding and activation functions. This post-translational modification, particularly by P/CAF, stimulates the activity of free E2F1, impacting cell cycle gene transcription.
Area of Science:
- Molecular Biology
- Cell Cycle Regulation
- Epigenetics
Background:
- The E2F-RB complex regulates cell cycle transcription, with RB repressing genes via histone deacetylase recruitment during G1.
- Phosphorylation of RB releases E2F, promoting S-phase gene transcription.
- E2F1 activity is modulated by acetylation (p300/CBP) and deacetylation (RB-associated deacetylase).
Purpose of the Study:
- To investigate whether E2F1 is subject to acetylation.
- To determine the functional consequences of E2F1 acetylation on its activity.
- To explore the interplay between acetylation and deacetylation in E2F1 regulation.
Main Methods:
- In vitro acetylation assays using E2F1 and acetyltransferases (P/CAF, p300/CBP).
- Detection of intracellular E2F1 acetylation.
- Analysis of E2F1 DNA-binding ability, activation potential, and protein half-life following acetylation.
- Assays to assess the effect of RB-associated histone deacetylase on E2F1.
Main Results:
- E2F1 is acetylated in vitro by P/CAF and p300/CBP, and also intracellularly.
- Acetylation sites are located near the E2F1 DNA-binding domain, involving conserved lysine residues.
- P/CAF-mediated acetylation increases E2F1's DNA-binding ability, activation potential, and protein half-life.
- RB-associated histone deacetylase can deacetylate E2F1.
Conclusions:
- Acetylation is a novel regulatory modification that enhances E2F1's transcriptional activation functions.
- Acetylation stimulates the activity of the non-RB bound, free form of E2F1.
- This acetylation process plays a significant role in regulating E2F1-dependent gene expression during the cell cycle.