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Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark
Published on: January 26, 2018
H4K20 methylation regulates quiescence and chromatin compaction
Adam G Evertts1, Amity L Manning, Xin Wang
1Department of Molecular Biology, Princeton University, Princeton, NJ 08544 Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129 Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104 Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, and Department of Biological Chemistry, David Geffen School of Medicine, Los Angeles, CA 90095.
Molecular Biology of the Cell
|August 9, 2013
Summary
Histone modifications regulate cell cycle exit. Increased H4K20me2 and H4K20me3 promote chromatin compaction and quiescence, while H4K20me1 is crucial for mitosis.
Area of Science:
- Cell Biology
- Epigenetics
- Molecular Biology
Background:
- Cellular quiescence involves complex regulatory networks.
- Chromatin structure and histone modifications are implicated in cell cycle control.
Purpose of the Study:
- To investigate the role of specific histone modifications in regulating the transition from cell proliferation to quiescence.
- To determine if changes in chromatin state directly influence cell cycle exit.
Main Methods:
- Mass spectrometry to analyze histone modifications in proliferating and quiescent human fibroblasts.
- Gene knockdown and overexpression of key enzymes involved in H4K20 methylation.
- Cell cycle phase analysis using flow cytometry.
Main Results:
- Quiescent fibroblasts exhibit increased chromatin compaction and elevated levels of H4K20me2 and H4K20me3.
- H4K20me1 levels increase post-S phase and are converted to H4K20me2/3 in quiescence.
- Knockdown of H4K20me3-producing enzyme impairs cell cycle exit and reduces chromatin compaction; Suv4-20h1 overexpression causes G2 arrest.
Conclusions:
- Histone H4K20 methylation states differentially regulate cell cycle progression and quiescence.
- H4K20me1 is important for mitosis, while H4K20me2/3 promote chromatin compaction and cell cycle exit into quiescence.
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