Cohesin Can Remain Associated with Chromosomes during DNA Replication
To ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created du...
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doaj-d2d6aa9273664aad9c878d52ce7838142020-11-25T01:55:15ZengElsevierCell Reports2211-12472017-09-0120122749275510.1016/j.celrep.2017.08.092Cohesin Can Remain Associated with Chromosomes during DNA ReplicationJames D.P. Rhodes0Judith H.I. Haarhuis1Jonathan B. Grimm2Benjamin D. Rowland3Luke D. Lavis4Kim A. Nasmyth5Department of Biochemistry, Oxford University, South Parks Road, Oxford, OX1 3QU, UKDepartment of Cell Biology, the Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the NetherlandsJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USADepartment of Cell Biology, the Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the NetherlandsJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USADepartment of Biochemistry, Oxford University, South Parks Road, Oxford, OX1 3QU, UKTo ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created during S phase is poorly understood, in particular whether they are derived from complexes that had entrapped DNAs prior to replication. To address this, we used selective photobleaching to determine whether cohesin associated with chromatin in G1 persists in situ after replication. We developed a non-fluorescent HaloTag ligand to discriminate the fluorescence recovery signal from labeling of newly synthesized Halo-tagged Scc1 protein (pulse-chase or pcFRAP). In cells where cohesin turnover is inactivated by deletion of WAPL, Scc1 can remain associated with chromatin throughout S phase. These findings suggest that cohesion might be generated by cohesin that is already bound to un-replicated DNA.http://www.sciencedirect.com/science/article/pii/S2211124717312342cohesincohesionestablishmentreplicationFRAPHaloTagTADs |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
James D.P. Rhodes Judith H.I. Haarhuis Jonathan B. Grimm Benjamin D. Rowland Luke D. Lavis Kim A. Nasmyth |
spellingShingle |
James D.P. Rhodes Judith H.I. Haarhuis Jonathan B. Grimm Benjamin D. Rowland Luke D. Lavis Kim A. Nasmyth Cohesin Can Remain Associated with Chromosomes during DNA Replication Cell Reports cohesin cohesion establishment replication FRAP HaloTag TADs |
author_facet |
James D.P. Rhodes Judith H.I. Haarhuis Jonathan B. Grimm Benjamin D. Rowland Luke D. Lavis Kim A. Nasmyth |
author_sort |
James D.P. Rhodes |
title |
Cohesin Can Remain Associated with Chromosomes during DNA Replication |
title_short |
Cohesin Can Remain Associated with Chromosomes during DNA Replication |
title_full |
Cohesin Can Remain Associated with Chromosomes during DNA Replication |
title_fullStr |
Cohesin Can Remain Associated with Chromosomes during DNA Replication |
title_full_unstemmed |
Cohesin Can Remain Associated with Chromosomes during DNA Replication |
title_sort |
cohesin can remain associated with chromosomes during dna replication |
publisher |
Elsevier |
series |
Cell Reports |
issn |
2211-1247 |
publishDate |
2017-09-01 |
description |
To ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created during S phase is poorly understood, in particular whether they are derived from complexes that had entrapped DNAs prior to replication. To address this, we used selective photobleaching to determine whether cohesin associated with chromatin in G1 persists in situ after replication. We developed a non-fluorescent HaloTag ligand to discriminate the fluorescence recovery signal from labeling of newly synthesized Halo-tagged Scc1 protein (pulse-chase or pcFRAP). In cells where cohesin turnover is inactivated by deletion of WAPL, Scc1 can remain associated with chromatin throughout S phase. These findings suggest that cohesion might be generated by cohesin that is already bound to un-replicated DNA. |
topic |
cohesin cohesion establishment replication FRAP HaloTag TADs |
url |
http://www.sciencedirect.com/science/article/pii/S2211124717312342 |
work_keys_str_mv |
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