Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts.
Interstrand crosslink (ICL)-inducing agents block the separation of the two DNA strands. They prevent transcription and replication and are used in clinics for the treatment of cancer and skin diseases. Here, we have introduced a single psoralen ICL at a specific site in plasmid DNA using a triplex-...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-04-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3078125?pdf=render |
| _version_ | 1817994849274560512 |
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| author | Cyrille Le Breton Magali Hennion Paola B Arimondo Olivier Hyrien |
| author_facet | Cyrille Le Breton Magali Hennion Paola B Arimondo Olivier Hyrien |
| author_sort | Cyrille Le Breton |
| collection | DOAJ |
| description | Interstrand crosslink (ICL)-inducing agents block the separation of the two DNA strands. They prevent transcription and replication and are used in clinics for the treatment of cancer and skin diseases. Here, we have introduced a single psoralen ICL at a specific site in plasmid DNA using a triplex-forming-oligonucleotide (TFO)-psoralen conjugate and studied its repair in Xenopus egg extracts that support nuclear assembly and replication of plasmid DNA. Replication forks arriving from either side stalled at the psoralen ICL. In contrast to previous observations with other ICL-inducing agents, the leading strands advanced up to the lesion without any prior pausing. Subsequently, incisions were introduced on one parental strand on both sides of the ICL. These incisions could be detected whether one or both forks reached the ICL. Using small molecule inhibitors, we found that the ATR-Chk1 pathway, but not the ATM-Chk2 pathway, stimulated both the incision step and the subsequent processing of the broken replication intermediates. Our results highlight both similarities and differences in fork stalling and repair induced by psoralen and by other ICL-forming agents. |
| first_indexed | 2024-04-14T01:58:29Z |
| format | Article |
| id | doaj.art-dcc7cf0c610a40d28740d5cfe95720e7 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T01:58:29Z |
| publishDate | 2011-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-dcc7cf0c610a40d28740d5cfe95720e72022-12-22T02:18:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-04-0164e1855410.1371/journal.pone.0018554Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts.Cyrille Le BretonMagali HennionPaola B ArimondoOlivier HyrienInterstrand crosslink (ICL)-inducing agents block the separation of the two DNA strands. They prevent transcription and replication and are used in clinics for the treatment of cancer and skin diseases. Here, we have introduced a single psoralen ICL at a specific site in plasmid DNA using a triplex-forming-oligonucleotide (TFO)-psoralen conjugate and studied its repair in Xenopus egg extracts that support nuclear assembly and replication of plasmid DNA. Replication forks arriving from either side stalled at the psoralen ICL. In contrast to previous observations with other ICL-inducing agents, the leading strands advanced up to the lesion without any prior pausing. Subsequently, incisions were introduced on one parental strand on both sides of the ICL. These incisions could be detected whether one or both forks reached the ICL. Using small molecule inhibitors, we found that the ATR-Chk1 pathway, but not the ATM-Chk2 pathway, stimulated both the incision step and the subsequent processing of the broken replication intermediates. Our results highlight both similarities and differences in fork stalling and repair induced by psoralen and by other ICL-forming agents.http://europepmc.org/articles/PMC3078125?pdf=render |
| spellingShingle | Cyrille Le Breton Magali Hennion Paola B Arimondo Olivier Hyrien Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. PLoS ONE |
| title | Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. |
| title_full | Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. |
| title_fullStr | Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. |
| title_full_unstemmed | Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. |
| title_short | Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts. |
| title_sort | replication fork stalling and processing at a single psoralen interstrand crosslink in xenopus egg extracts |
| url | http://europepmc.org/articles/PMC3078125?pdf=render |
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