A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint
DNA damage checkpoints arrest cell cycle progression to facilitate DNA repair. The ability to survive genotoxic insults depends not only on the initiation of cell cycle checkpoints but also on checkpoint maintenance. While activation of DNA damage checkpoints has been studied extensively, molecular...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2010
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| Online Access: | http://hdl.handle.net/1721.1/55355 https://orcid.org/0000-0002-3202-4678 https://orcid.org/0000-0002-9547-3251 |
| _version_ | 1811096204217417728 |
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| author | Yaffe, Michael B. Brummelkamp, Thijn R. Reinhardt, Hans Ostheimer, Gerard Gardino, Alexandra Kate van Vugt, Marcel A. T. M. Smerdon, Stephen J. Carr, Steven A. Lewis, Timothy A. Pawson, Tony Li, Jeijin Keezer, Susan M. Miao, Hua Tan, Chris S. Ong, Shao-En Linding, Rune |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Yaffe, Michael B. Brummelkamp, Thijn R. Reinhardt, Hans Ostheimer, Gerard Gardino, Alexandra Kate van Vugt, Marcel A. T. M. Smerdon, Stephen J. Carr, Steven A. Lewis, Timothy A. Pawson, Tony Li, Jeijin Keezer, Susan M. Miao, Hua Tan, Chris S. Ong, Shao-En Linding, Rune |
| author_sort | Yaffe, Michael B. |
| collection | MIT |
| description | DNA damage checkpoints arrest cell cycle progression to facilitate DNA repair. The ability to survive genotoxic insults depends not only on the initiation of cell cycle checkpoints but also on checkpoint maintenance. While activation of DNA damage checkpoints has been studied extensively, molecular mechanisms involved in sustaining and ultimately inactivating cell cycle checkpoints are largely unknown. Here, we explored feedback mechanisms that control the maintenance and termination of checkpoint function by computationally identifying an evolutionary conserved mitotic phosphorylation network within the DNA damage response. We demonstrate that the non-enzymatic checkpoint adaptor protein 53BP1 is an in vivo target of the cell cycle kinases Cyclin-dependent kinase-1 and Polo-like kinase-1 (Plk1). We show that Plk1 binds 53BP1 during mitosis and that this interaction is required for proper inactivation of the DNA damage checkpoint. 53BP1 mutants that are unable to bind Plk1 fail to restart the cell cycle after ionizing radiation-mediated cell cycle arrest. Importantly, we show that Plk1 also phosphorylates the 53BP1-binding checkpoint kinase Chk2 to inactivate its FHA domain and inhibit its kinase activity in mammalian cells. Thus, a mitotic kinase-mediated negative feedback loop regulates the ATM-Chk2 branch of the DNA damage signaling network by phosphorylating conserved sites in 53BP1 and Chk2 to inactivate checkpoint signaling and control checkpoint duration. |
| first_indexed | 2024-09-23T16:40:14Z |
| format | Article |
| id | mit-1721.1/55355 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:40:14Z |
| publishDate | 2010 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/553552022-09-29T20:39:43Z A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint Yaffe, Michael B. Brummelkamp, Thijn R. Reinhardt, Hans Ostheimer, Gerard Gardino, Alexandra Kate van Vugt, Marcel A. T. M. Smerdon, Stephen J. Carr, Steven A. Lewis, Timothy A. Pawson, Tony Li, Jeijin Keezer, Susan M. Miao, Hua Tan, Chris S. Ong, Shao-En Linding, Rune Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Koch Institute for Integrative Cancer Research at MIT Yaffe, Michael B. Yaffe, Michael B. Brummelkamp, Thijn R. Reinhardt, Hans Ostheimer, Gerard Gardino, Alexandra Kate van Vugt, Marcel A. T. M. DNA damage checkpoints arrest cell cycle progression to facilitate DNA repair. The ability to survive genotoxic insults depends not only on the initiation of cell cycle checkpoints but also on checkpoint maintenance. While activation of DNA damage checkpoints has been studied extensively, molecular mechanisms involved in sustaining and ultimately inactivating cell cycle checkpoints are largely unknown. Here, we explored feedback mechanisms that control the maintenance and termination of checkpoint function by computationally identifying an evolutionary conserved mitotic phosphorylation network within the DNA damage response. We demonstrate that the non-enzymatic checkpoint adaptor protein 53BP1 is an in vivo target of the cell cycle kinases Cyclin-dependent kinase-1 and Polo-like kinase-1 (Plk1). We show that Plk1 binds 53BP1 during mitosis and that this interaction is required for proper inactivation of the DNA damage checkpoint. 53BP1 mutants that are unable to bind Plk1 fail to restart the cell cycle after ionizing radiation-mediated cell cycle arrest. Importantly, we show that Plk1 also phosphorylates the 53BP1-binding checkpoint kinase Chk2 to inactivate its FHA domain and inhibit its kinase activity in mammalian cells. Thus, a mitotic kinase-mediated negative feedback loop regulates the ATM-Chk2 branch of the DNA damage signaling network by phosphorylating conserved sites in 53BP1 and Chk2 to inactivate checkpoint signaling and control checkpoint duration. National Institutes of Health (contract no. N01-CO-12400) National Cancer Institute’s Initiative for Chemical Genetics 2010-05-28T20:09:02Z 2010-05-28T20:09:02Z 2010-01 2010-12 Article http://purl.org/eprint/type/JournalArticle 1544-9173 1545-7885 http://hdl.handle.net/1721.1/55355 van Vugt MATM, Gardino AK, Linding R, Ostheimer GJ, Reinhardt HC, et al. (2010) A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint. PLoS Biol 8(1): e1000287. doi:10.1371/journal.pbio.1000287 https://orcid.org/0000-0002-3202-4678 https://orcid.org/0000-0002-9547-3251 en_US http://dx.doi.org/10.1371/journal.pbio.1000287 PLoS Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Public Library of Science PLoS |
| spellingShingle | Yaffe, Michael B. Brummelkamp, Thijn R. Reinhardt, Hans Ostheimer, Gerard Gardino, Alexandra Kate van Vugt, Marcel A. T. M. Smerdon, Stephen J. Carr, Steven A. Lewis, Timothy A. Pawson, Tony Li, Jeijin Keezer, Susan M. Miao, Hua Tan, Chris S. Ong, Shao-En Linding, Rune A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title | A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title_full | A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title_fullStr | A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title_full_unstemmed | A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title_short | A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint |
| title_sort | mitotic phosphorylation feedback network connects cdk1 plk1 53bp1 and chk2 to inactivate the g2 m dna damage checkpoint |
| url | http://hdl.handle.net/1721.1/55355 https://orcid.org/0000-0002-3202-4678 https://orcid.org/0000-0002-9547-3251 |
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