Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B
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-...
| Main Authors: | , |
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| Format: | Journal article |
| Language: | English |
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Rockefeller University Press
2001
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| Subjects: |
| _version_ | 1826284177409966080 |
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| author | Kimura, H Cook, P |
| author_facet | Kimura, H Cook, P |
| author_sort | Kimura, H |
| collection | OXFORD |
| description | 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 neterokaryons, photobleaching revealed that H2b-GFP exchanged more rapidly than H3-GFP. About 3% of H2B exchanged within minutes, whereas ~40% did so slowly (t1/2 ~ 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. |
| first_indexed | 2024-03-07T01:09:58Z |
| format | Journal article |
| id | oxford-uuid:8ca773da-ccd1-4e93-ad35-c9facee8be8f |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:09:58Z |
| publishDate | 2001 |
| publisher | Rockefeller University Press |
| record_format | dspace |
| spelling | oxford-uuid:8ca773da-ccd1-4e93-ad35-c9facee8be8f2022-03-26T22:45:58ZKinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2BJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:8ca773da-ccd1-4e93-ad35-c9facee8be8fPathologyEnglishOxford University Research Archive - ValetRockefeller University Press2001Kimura, HCook, PHistones 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 neterokaryons, photobleaching revealed that H2b-GFP exchanged more rapidly than H3-GFP. About 3% of H2B exchanged within minutes, whereas ~40% did so slowly (t1/2 ~ 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. |
| spellingShingle | Pathology Kimura, H Cook, P Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title | Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title_full | Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title_fullStr | Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title_full_unstemmed | Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title_short | Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B |
| title_sort | kinetics of core histones in living human cells little exchange of h3 and h4 and some rapid exchange of h2b |
| topic | Pathology |
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