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...
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| Format: | Article |
| Language: | English |
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Wiley
2010-01-01
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| Series: | Journal of Nucleic Acids |
| Online Access: | http://dx.doi.org/10.4061/2010/319142 |
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| author | Daniël O. Warmerdam Roland Kanaar Veronique A. J. Smits |
| author_facet | Daniël O. Warmerdam Roland Kanaar Veronique A. J. Smits |
| author_sort | Daniël O. Warmerdam |
| collection | DOAJ |
| 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 | Article |
| id | doaj-art-ef0e6ee70cbb48489832c4de20e48bd6 |
| institution | Kabale University |
| issn | 2090-021X |
| language | English |
| publishDate | 2010-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nucleic Acids |
| spelling | doaj-art-ef0e6ee70cbb48489832c4de20e48bd62025-02-03T01:23:51ZengWileyJournal of Nucleic Acids2090-021X2010-01-01201010.4061/2010/319142319142Differential Dynamics of ATR-Mediated Checkpoint RegulatorsDaniël O. Warmerdam0Roland Kanaar1Veronique A. J. Smits2Department of Cell Biology and Genetics, Cancer Genome Center, Erasmus MC, Dr. Molewaterplein 50, 3015 GE Rotterdam, The NetherlandsDepartment of Cell Biology and Genetics, Cancer Genome Center, Erasmus MC, Dr. Molewaterplein 50, 3015 GE Rotterdam, The NetherlandsUnidad de Investigación, Hospital Universitario de Canarias, Ofra s/n, La Laguna 38320, Tenerife, SpainThe 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.http://dx.doi.org/10.4061/2010/319142 |
| spellingShingle | Daniël O. Warmerdam Roland Kanaar Veronique A. J. Smits Differential Dynamics of ATR-Mediated Checkpoint Regulators Journal of Nucleic Acids |
| 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 |
| url | http://dx.doi.org/10.4061/2010/319142 |
| work_keys_str_mv | AT danielowarmerdam differentialdynamicsofatrmediatedcheckpointregulators AT rolandkanaar differentialdynamicsofatrmediatedcheckpointregulators AT veroniqueajsmits differentialdynamicsofatrmediatedcheckpointregulators |