Kinetics of Core Histones in Living Human Cells

By H. Kimura, Peter R. Cook. Journal of Cell Biology, 2001

733 citations recorded by the source corpus.

Histones H2A and H2B form part of the same nucleosomal structure as H3 and H4. Stable HeLa cell lines expressing histones H2B, H3, and H4 tagged with green fluorescent protein (GFP) were established; the tagged molecules were assembled into nucleosomes. Although H2B-GFP was distributed like DNA, H3-GFP and H4-GFP were concentrated in euchromatin during interphase and in R-bands in mitotic chromosomes. These differences probably result from an unregulated production of tagged histones and differences in exchange. In both single cells and heterokaryons, photobleaching revealed that H2B-GFP exchanged more rapidly than H3-GFP and H4-GFP. About 3% of H2B exchanged within minutes, whereas ∼40% did so slowly (t 1/2 ∼ 130 min). The rapidly exchanging fraction disappeared in 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole and so may represent H2B in transcriptionally active chromatin. The slowly exchanging fraction was probably associated with chromatin domains surrounding active units. H3-GFP and H4-GFP were assembled into chromatin when DNA was replicated, and then >80% remained bound permanently. These results reveal that the inner core of the nucleosome is very stable, whereas H2B on the surface of active nucleosomes exchanges continually.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/21a72a6834c0c19316ee171363fe6ba6cae35d86

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Citations in the corpus

A histone H3 methyltransferase controls DNA methylation in Neurospora crassa (2001, unknown): “Of note, histones H3 and H4 remain tightly bound to DNA in vivo , unlike histones H2A and H2”
Epigenetic inheritance during the cell cycle (2009, background): “1 and histone H4 is minimal compared with the rapid exchange of H2A and H2”
The γ‐H2A.X: Is it just a surrogate marker of double‐strand breaks or much more? (2008, background): “In favor of this model, slow exchange in chromatin was detected earlier in experiments with GFP-tagged histones H2B, H2A, H3, and H4 [Kimura and Cook, 2001; Gautier et al., 2004].”
From silencing to gene expression: real-time analysis in single cells. (2004, background): “…can be activated at this array in an experi- been shown to be tightly associated with chromatin mentally regulated manner, we were able to study in (Kimura and Cook, 2001), it will be of particular interest living cells the dynamic changes that occur as hetero- to characterize the histone…”
Hyperdynamic plasticity of chromatin proteins in pluripotent embryonic stem cells. (2006, background): “, 2000), while the recovery time of core histones has been shown to be on the order of hours (Kimura and Cook, 2001; Phair et al., 2004).”
The histone variant H3.3 marks active chromatin by replication-independent nucleosome assembly. (2002, background): “these histones (Wolffe, 1998), and the proper behavior of GFP-tagged histones has been extensively documented (Kimura and Cook, 2001).”
Chromatin challenges during DNA replication and repair. (2007, unknown): “H3-H4 Dynamics at Repair Sites Within chromatin the overall exchange of H2A-H2B is rapid in comparison to that of H3-H4 (Kimura and Cook, 2001).”
The role of chromatin during transcription. (2007, background): “In addition, a cytological study found that histone H2B rapidly exchanged in and out of a nucleosome relative to H3 and H4 (Kimura and Cook, 2001), which suggests that histone displacement does occur in vivo.”