4D Visualization of replication foci in mammalian cells corresponding to individual replicons
Since the pioneering proposal of the replicon model of DNA replication 50 years ago, the predicted replicons have not been identified and quantified at the cellular level. Here, we combine conventional and super-resolution microscopy of replication sites in live and fixed cells with computational im...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Journal article |
| Published: |
Nature Publishing Group
2016
|
| _version_ | 1797068752001433600 |
|---|---|
| author | Chagin, V Casas-Delucchi, C Reinhart, MR Schermelleh, L Markaki, Y Maiser, A Bolius, J Bensimon, AB Filies, M Domaing, P Rozanov, Y Leonhardt, H Cardoso, M |
| author_facet | Chagin, V Casas-Delucchi, C Reinhart, MR Schermelleh, L Markaki, Y Maiser, A Bolius, J Bensimon, AB Filies, M Domaing, P Rozanov, Y Leonhardt, H Cardoso, M |
| author_sort | Chagin, V |
| collection | OXFORD |
| description | Since the pioneering proposal of the replicon model of DNA replication 50 years ago, the predicted replicons have not been identified and quantified at the cellular level. Here, we combine conventional and super-resolution microscopy of replication sites in live and fixed cells with computational image analysis. We complement these data with genome size measurements, comprehensive analysis of S-phase dynamics and quantification of replication fork speed and replicon size in human and mouse cells. These multidimensional analyses demonstrate that replication foci (RFi) in three-dimensional (3D) preserved somatic mammalian cells can be optically resolved down to single replicons throughout S-phase. This challenges the conventional interpretation of nuclear RFi as replication factories, that is, the complex entities that process multiple clustered replicons. Accordingly, 3D genome organization and duplication can be now followed within the chromatin context at the level of individual replicons. |
| first_indexed | 2024-03-06T22:14:38Z |
| format | Journal article |
| id | oxford-uuid:52fa8365-00c6-4761-9b40-3ad3b608b4b1 |
| institution | University of Oxford |
| last_indexed | 2024-03-06T22:14:38Z |
| publishDate | 2016 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | oxford-uuid:52fa8365-00c6-4761-9b40-3ad3b608b4b12022-03-26T16:28:49Z4D Visualization of replication foci in mammalian cells corresponding to individual repliconsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:52fa8365-00c6-4761-9b40-3ad3b608b4b1Symplectic Elements at OxfordNature Publishing Group2016Chagin, VCasas-Delucchi, CReinhart, MRSchermelleh, LMarkaki, YMaiser, ABolius, JBensimon, ABFilies, MDomaing, PRozanov, YLeonhardt, HCardoso, MSince the pioneering proposal of the replicon model of DNA replication 50 years ago, the predicted replicons have not been identified and quantified at the cellular level. Here, we combine conventional and super-resolution microscopy of replication sites in live and fixed cells with computational image analysis. We complement these data with genome size measurements, comprehensive analysis of S-phase dynamics and quantification of replication fork speed and replicon size in human and mouse cells. These multidimensional analyses demonstrate that replication foci (RFi) in three-dimensional (3D) preserved somatic mammalian cells can be optically resolved down to single replicons throughout S-phase. This challenges the conventional interpretation of nuclear RFi as replication factories, that is, the complex entities that process multiple clustered replicons. Accordingly, 3D genome organization and duplication can be now followed within the chromatin context at the level of individual replicons. |
| spellingShingle | Chagin, V Casas-Delucchi, C Reinhart, MR Schermelleh, L Markaki, Y Maiser, A Bolius, J Bensimon, AB Filies, M Domaing, P Rozanov, Y Leonhardt, H Cardoso, M 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title | 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title_full | 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title_fullStr | 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title_full_unstemmed | 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title_short | 4D Visualization of replication foci in mammalian cells corresponding to individual replicons |
| title_sort | 4d visualization of replication foci in mammalian cells corresponding to individual replicons |
| work_keys_str_mv | AT chaginv 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT casasdelucchic 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT reinhartmr 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT schermellehl 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT markakiy 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT maisera 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT boliusj 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT bensimonab 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT filiesm 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT domaingp 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT rozanovy 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT leonhardth 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons AT cardosom 4dvisualizationofreplicationfociinmammaliancellscorrespondingtoindividualreplicons |