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Differential Dynamics of ATR-Mediated Checkpoint Regulators
The ATR-Chk1 checkpoint pathway is activated by UV-induced DNA lesions and replication stress. Little was known about the spatio and temporal behaviour of the proteins involved, and we, therefore, examined the behaviour of the ATRIP-ATR and Rad9-Rad1-Hus1 putative DNA damage sensor complexes and the...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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SAGE-Hindawi Access to Research
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2933903/ https://www.ncbi.nlm.nih.gov/pubmed/20847938 http://dx.doi.org/10.4061/2010/319142 |
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author | Warmerdam, Daniël O. Kanaar, Roland Smits, Veronique A. J. |
author_facet | Warmerdam, Daniël O. Kanaar, Roland Smits, Veronique A. J. |
author_sort | Warmerdam, Daniël O. |
collection | PubMed |
description | The ATR-Chk1 checkpoint pathway is activated by UV-induced DNA lesions and replication stress. Little was known about the spatio and temporal behaviour of the proteins involved, and we, therefore, examined the behaviour of the ATRIP-ATR and Rad9-Rad1-Hus1 putative DNA damage sensor complexes and the downstream effector kinase Chk1. We developed assays for the generation and validation of stable cell lines expressing GFP-fusion proteins. Photobleaching experiments in living cells expressing these fusions indicated that after UV-induced DNA damage, ATRIP associates more transiently with damaged chromatin than members of the Rad9-Rad1-Hus1 complex. Interestingly, ATRIP directly associated with locally induced UV damage, whereas Rad9 bound in a cooperative manner, which can be explained by the Rad17-dependent loading of Rad9 onto damaged chromatin. Although Chk1 dissociates from the chromatin upon UV damage, no change in the mobility of GFP-Chk1 was observed, supporting the notion that Chk1 is a highly dynamic protein. |
format | Text |
id | pubmed-2933903 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | SAGE-Hindawi Access to Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-29339032010-09-16 Differential Dynamics of ATR-Mediated Checkpoint Regulators Warmerdam, Daniël O. Kanaar, Roland Smits, Veronique A. J. J Nucleic Acids Research Article The ATR-Chk1 checkpoint pathway is activated by UV-induced DNA lesions and replication stress. Little was known about the spatio and temporal behaviour of the proteins involved, and we, therefore, examined the behaviour of the ATRIP-ATR and Rad9-Rad1-Hus1 putative DNA damage sensor complexes and the downstream effector kinase Chk1. We developed assays for the generation and validation of stable cell lines expressing GFP-fusion proteins. Photobleaching experiments in living cells expressing these fusions indicated that after UV-induced DNA damage, ATRIP associates more transiently with damaged chromatin than members of the Rad9-Rad1-Hus1 complex. Interestingly, ATRIP directly associated with locally induced UV damage, whereas Rad9 bound in a cooperative manner, which can be explained by the Rad17-dependent loading of Rad9 onto damaged chromatin. Although Chk1 dissociates from the chromatin upon UV damage, no change in the mobility of GFP-Chk1 was observed, supporting the notion that Chk1 is a highly dynamic protein. SAGE-Hindawi Access to Research 2010-08-17 /pmc/articles/PMC2933903/ /pubmed/20847938 http://dx.doi.org/10.4061/2010/319142 Text en Copyright © 2010 Daniël O. Warmerdam et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Warmerdam, Daniël O. Kanaar, Roland Smits, Veronique A. J. Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title | Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title_full | Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title_fullStr | Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title_full_unstemmed | Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title_short | Differential Dynamics of ATR-Mediated Checkpoint Regulators |
title_sort | differential dynamics of atr-mediated checkpoint regulators |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2933903/ https://www.ncbi.nlm.nih.gov/pubmed/20847938 http://dx.doi.org/10.4061/2010/319142 |
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