Artemis Regulates Cell Cycle Recovery from the S Phase Checkpoint by Promoting Degradation of Cyclin E
Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser516 and Ser645) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G2/M checkpoint in respo...
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Published in | The Journal of biological chemistry Vol. 284; no. 27; pp. 18236 - 18243 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
03.07.2009
American Society for Biochemistry and Molecular Biology |
Subjects | |
Online Access | Get full text |
ISSN | 0021-9258 1083-351X 1083-351X |
DOI | 10.1074/jbc.M109.002584 |
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Abstract | Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser516 and Ser645) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G2/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser516 and Ser645 residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCFFbw7 E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser516 and Ser645 sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. |
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AbstractList | Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser(516) and Ser(645)) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G(2)/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser(516) and Ser(645) residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCF(Fbw7) E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser(516) and Ser(645) sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser(516) and Ser(645)) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G(2)/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser(516) and Ser(645) residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCF(Fbw7) E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser(516) and Ser(645) sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E.Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser(516) and Ser(645)) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G(2)/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser(516) and Ser(645) residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCF(Fbw7) E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser(516) and Ser(645) sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser 516 and Ser 645 ) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G 2 /M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser 516 and Ser 645 residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCF Fbw7 E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser 516 and Ser 645 sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser516 and Ser645) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G2/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser516 and Ser645 residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCFFbw7 E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser516 and Ser645 sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser 516 and Ser 645 ) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G 2 /M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser 516 and Ser 645 residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCF Fbw7 E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser 516 and Ser 645 sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser⁵¹³ and Ser³´⁵) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G(2)/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser⁵¹³ and Ser³´⁵ residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCFFbw· E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser⁵¹³ and Ser³´⁵ sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in Artemis (Ser⁵¹⁶ and Ser⁶⁴⁵) that are subject to phosphorylation by ATM and are involved in mediating recovery from the G₂/M checkpoint in response to ionizing radiation. Here we show that these same sites are also phosphorylated by ATR in response to various types of replication stress including UVC, aphidicolin, and hydroxyurea. We also show that mutation of the Ser⁵¹⁶ and Ser⁶⁴⁵ residues causes a prolonged S phase checkpoint recovery after treatment with UV or aphidicolin, and that this delayed recovery process coincides with a prolonged stabilization of cyclin E and down-regulation of Cdk2 kinase activity. Furthermore, we show that Artemis interacts with the F-box protein Fbw7, and that this interaction regulates cyclin E degradation through the SCFFbw⁷ E3 ubiquitin ligase complex. The interaction between Artemis and Fbw7 is regulated by phosphorylation of Ser⁵¹⁶ and Ser⁶⁴⁵ sites that occur in response to replication stress. Thus, our findings suggest a novel pathway of recovery from the S phase checkpoint in that in response to replication stress phosphorylation of Artemis by ATR enhances its interaction with Fbw7, which in turn promotes ubiquitylation and the ultimate degradation of cyclin E. |
Author | Legerski, Randy J. Geng, Liyi Teng, Lisong Zhang, Xiaoshan Wang, Haiyong |
Author_xml | – sequence: 1 givenname: Haiyong surname: Wang fullname: Wang, Haiyong organization: Department of Oncology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China – sequence: 2 givenname: Xiaoshan surname: Zhang fullname: Zhang, Xiaoshan organization: Department of Genetics, University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030 – sequence: 3 givenname: Liyi surname: Geng fullname: Geng, Liyi organization: Department of Genetics, University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030 – sequence: 4 givenname: Lisong surname: Teng fullname: Teng, Lisong organization: Department of Oncology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China – sequence: 5 givenname: Randy J. surname: Legerski fullname: Legerski, Randy J. email: rlegersk@mdanderson.org organization: Department of Genetics, University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030 |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/19423708$$D View this record in MEDLINE/PubMed |
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Snippet | Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in... Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in... Artemis, a member of the SNM1 gene family, is a known phosphorylation target of ATM, ATR, and DNA-PKcs. We have previously identified two serine residues in... |
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StartPage | 18236 |
SubjectTerms | Ataxia Telangiectasia Mutated Proteins Cell Cycle Proteins - metabolism Cell Line Cyclin E - metabolism Cyclin-Dependent Kinase 2 - metabolism DNA-Binding Proteins DNA: , Repair, Recombination, and Chromosome Dynamics Endonucleases F-Box Proteins - metabolism F-Box-WD Repeat-Containing Protein 7 Flow Cytometry Humans Kidney - cytology Mutagenesis, Site-Directed Nuclear Proteins - genetics Nuclear Proteins - metabolism Oncogene Proteins - metabolism Phosphorylation - physiology Protein-Serine-Threonine Kinases - metabolism RNA, Small Interfering S Phase - physiology Serine - metabolism Stress, Physiological - physiology Ubiquitin - metabolism Ubiquitin-Protein Ligases - metabolism |
Title | Artemis Regulates Cell Cycle Recovery from the S Phase Checkpoint by Promoting Degradation of Cyclin E |
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