MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach
14-3-3 proteins play critical roles in controlling multiple aspects of the cellular response to stress and DNA damage including regulation of metabolism, cell cycle progression, cell migration, and apoptotic cell death by binding to protein substrates of basophilic protein kinases following their ph...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-04-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1148933/full |
| _version_ | 1827971545759219712 |
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| author | Justine R. Stehn Scott R. Floyd Erik W. Wilker H. Christian Reinhardt Scott M. Clarke Qiuying Huang Roberto D. Polakiewicz Nahum Sonenberg Yi Wen Kong Yi Wen Kong Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe |
| author_facet | Justine R. Stehn Scott R. Floyd Erik W. Wilker H. Christian Reinhardt Scott M. Clarke Qiuying Huang Roberto D. Polakiewicz Nahum Sonenberg Yi Wen Kong Yi Wen Kong Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe |
| author_sort | Justine R. Stehn |
| collection | DOAJ |
| description | 14-3-3 proteins play critical roles in controlling multiple aspects of the cellular response to stress and DNA damage including regulation of metabolism, cell cycle progression, cell migration, and apoptotic cell death by binding to protein substrates of basophilic protein kinases following their phosphorylation on specific serine/threonine residues. Although over 200 mammalian proteins that bind to 14-3-3 have been identified, largely through proteomic studies, in many cases the relevant protein kinase responsible for conferring 14-3-3-binding to these proteins is not known. To facilitate the identification of kinase-specific 14-3-3 clients, we developed a biochemical approach using high-density protein filter arrays and identified the translational regulatory molecule PABPC1 as a substrate for Chk1 and MAPKAP Kinase-2 (MK2) in vitro, and for MK2 in vivo, whose phosphorylation results in 14-3-3-binding. We identify Ser-470 on PABPC1 within the linker region connecting the RRM domains to the PABC domain as the critical 14-3-3-binding site, and demonstrate that loss of PABPC1 binding to 14-3-3 results in increased cell proliferation and decreased cell death in response to UV-induced DNA damage. |
| first_indexed | 2024-04-09T19:10:25Z |
| format | Article |
| id | doaj.art-a76d35ddc948487f896db3bbbe2880e5 |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-09T19:10:25Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-a76d35ddc948487f896db3bbbe2880e52023-04-06T16:23:55ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2023-04-011010.3389/fmolb.2023.11489331148933MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approachJustine R. Stehn0Scott R. Floyd1Erik W. Wilker2H. Christian Reinhardt3Scott M. Clarke4Qiuying Huang5Roberto D. Polakiewicz6Nahum Sonenberg7Yi Wen Kong8Yi Wen Kong9Michael B. Yaffe10Michael B. Yaffe11Michael B. Yaffe12Michael B. Yaffe13Michael B. Yaffe14Michael B. Yaffe15David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesCell Signaling Technology, Danvers, MA, United StatesRosalind and Morris Goodman Cancer Centre, Department of Biochemistry, McGill University, Montreal, QC, CanadaDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesCenter for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, MA, United StatesDavid H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United StatesCenter for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, MA, United StatesDepartment of Biology, Massachusetts Institute of Technology, Cambridge, MA, United StatesDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United StatesDivisions of Surgical Oncology, Trauma, and Surgical Critical Care, Beth Israel Deaconess Medical Center, Department of Surgery, Harvard Medical School, Boston, MA, United StatesSurgical Oncology Program, National Cancer Institute, National Institutes of Health, Bethesda, MD, United States14-3-3 proteins play critical roles in controlling multiple aspects of the cellular response to stress and DNA damage including regulation of metabolism, cell cycle progression, cell migration, and apoptotic cell death by binding to protein substrates of basophilic protein kinases following their phosphorylation on specific serine/threonine residues. Although over 200 mammalian proteins that bind to 14-3-3 have been identified, largely through proteomic studies, in many cases the relevant protein kinase responsible for conferring 14-3-3-binding to these proteins is not known. To facilitate the identification of kinase-specific 14-3-3 clients, we developed a biochemical approach using high-density protein filter arrays and identified the translational regulatory molecule PABPC1 as a substrate for Chk1 and MAPKAP Kinase-2 (MK2) in vitro, and for MK2 in vivo, whose phosphorylation results in 14-3-3-binding. We identify Ser-470 on PABPC1 within the linker region connecting the RRM domains to the PABC domain as the critical 14-3-3-binding site, and demonstrate that loss of PABPC1 binding to 14-3-3 results in increased cell proliferation and decreased cell death in response to UV-induced DNA damage.https://www.frontiersin.org/articles/10.3389/fmolb.2023.1148933/full14-3-3polyA-binding proteinMAPKAP Kinase-2signal transductionDNA damage |
| spellingShingle | Justine R. Stehn Scott R. Floyd Erik W. Wilker H. Christian Reinhardt Scott M. Clarke Qiuying Huang Roberto D. Polakiewicz Nahum Sonenberg Yi Wen Kong Yi Wen Kong Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe Michael B. Yaffe MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach Frontiers in Molecular Biosciences 14-3-3 polyA-binding protein MAPKAP Kinase-2 signal transduction DNA damage |
| title | MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach |
| title_full | MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach |
| title_fullStr | MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach |
| title_full_unstemmed | MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach |
| title_short | MAPKAP Kinase-2 phosphorylation of PABPC1 controls its interaction with 14-3-3 proteins after DNA damage: A combined kinase and protein array approach |
| title_sort | mapkap kinase 2 phosphorylation of pabpc1 controls its interaction with 14 3 3 proteins after dna damage a combined kinase and protein array approach |
| topic | 14-3-3 polyA-binding protein MAPKAP Kinase-2 signal transduction DNA damage |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1148933/full |
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