CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response

DNA damage triggers a phosphorylation-based signaling cascade known as the DNA damage response. p19INK4d, a member of the INK4 family of CDK4/6 inhibitors, has been reported to participate in the DNA damage response promoting DNA repair and cell survival. Here, we provide mechanistic insight into th...

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Autores principales:	Marazita, Mariela C., Ogara, Maria Florencia, Sonzogni, Silvina Verónica, Martí, Marcelo Adrián, Dusetti, Nelson J., Pignataro, Omar Pedro, Cánepa, Eduardo Tomás
Publicado:	2012
Materias:	amyloid beta protein ATM protein ATR protein checkpoint kinase 1 checkpoint kinase 2 cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase 4 cyclin dependent kinase 6 cyclin dependent kinase inhibitor 2D phosphate p 32 serine threonine CDK2 protein, human DNA article cell nucleus cell survival conformational transition controlled study DNA damage DNA repair point mutation protein phosphorylation signal transduction ultraviolet radiation cell cycle cell line cell strain HEK293 drug effect gene expression regulation genetics human metabolism mutation phosphorylation protein transport radiation exposure Amyloid beta-Peptides Cell Cycle Cell Line Cell Survival Cisplatin Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinase Inhibitor p19 DNA Damage DNA Repair Gene Expression Regulation HEK293 Cells Humans Mutation Phosphorylation Protein Transport Signal Transduction Ultraviolet Rays
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v7_n4_p_Marazita http://hdl.handle.net/20.500.12110/paper_19326203_v7_n4_p_Marazita
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_19326203_v7_n4_p_Marazita
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spelling	paper:paper_19326203_v7_n4_p_Marazita2023-06-08T16:31:05Z CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response Marazita, Mariela C. Ogara, Maria Florencia Sonzogni, Silvina Verónica Martí, Marcelo Adrián Dusetti, Nelson J. Pignataro, Omar Pedro Cánepa, Eduardo Tomás amyloid beta protein ATM protein ATR protein checkpoint kinase 1 checkpoint kinase 2 cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase 4 cyclin dependent kinase 6 cyclin dependent kinase inhibitor 2D phosphate p 32 serine threonine amyloid beta protein CDK2 protein, human cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase inhibitor 2D DNA article cell nucleus cell survival conformational transition controlled study DNA damage DNA repair point mutation protein phosphorylation signal transduction ultraviolet radiation cell cycle cell line cell strain HEK293 DNA damage DNA repair drug effect gene expression regulation genetics human metabolism mutation phosphorylation protein transport radiation exposure Amyloid beta-Peptides Cell Cycle Cell Line Cell Survival Cisplatin Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinase Inhibitor p19 DNA DNA Damage DNA Repair Gene Expression Regulation HEK293 Cells Humans Mutation Phosphorylation Protein Transport Signal Transduction Ultraviolet Rays DNA damage triggers a phosphorylation-based signaling cascade known as the DNA damage response. p19INK4d, a member of the INK4 family of CDK4/6 inhibitors, has been reported to participate in the DNA damage response promoting DNA repair and cell survival. Here, we provide mechanistic insight into the activation mechanism of p19INK4d linked to the response to DNA damage. Results showed that p19INK4d becomes phosphorylated following UV radiation, b-amyloid peptide and cisplatin treatments. ATM-Chk2/ATR-Chk1 signaling pathways were found to be differentially involved in p19INK4d phosphorylation depending on the type of DNA damage. Two sequential phosphorylation events at serine 76 and threonine 141 were identified using p19INK4d single-point mutants in metabolic labeling assays with 32P-orthophosphate. CDK2 and PKA were found to participate in p19INK4d phosphorylation process and that they would mediate serine 76 and threonine 141 modifications respectively. Nuclear translocation of p19INK4d induced by DNA damage was shown to be dependent on serine 76 phosphorylation. Most importantly, both phosphorylation sites were found to be crucial for p19INK4d function in DNA repair and cell survival. In contrast, serine 76 and threonine 141 were dispensable for CDK4/6 inhibition highlighting the independence of p19INK4d functions, in agreement with our previous findings. These results constitute the first description of the activation mechanism of p19INK4d in response to genotoxic stress and demonstrate the functional relevance of this activation following DNA damage. © 2012 Marazita et al. Fil:Marazita, M.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Florencia Ogara, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Sonzogni, S.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Martí, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Dusetti, N.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pignataro, O.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cánepa, E.T. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2012 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v7_n4_p_Marazita http://hdl.handle.net/20.500.12110/paper_19326203_v7_n4_p_Marazita
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	amyloid beta protein ATM protein ATR protein checkpoint kinase 1 checkpoint kinase 2 cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase 4 cyclin dependent kinase 6 cyclin dependent kinase inhibitor 2D phosphate p 32 serine threonine amyloid beta protein CDK2 protein, human cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase inhibitor 2D DNA article cell nucleus cell survival conformational transition controlled study DNA damage DNA repair point mutation protein phosphorylation signal transduction ultraviolet radiation cell cycle cell line cell strain HEK293 DNA damage DNA repair drug effect gene expression regulation genetics human metabolism mutation phosphorylation protein transport radiation exposure Amyloid beta-Peptides Cell Cycle Cell Line Cell Survival Cisplatin Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinase Inhibitor p19 DNA DNA Damage DNA Repair Gene Expression Regulation HEK293 Cells Humans Mutation Phosphorylation Protein Transport Signal Transduction Ultraviolet Rays
spellingShingle	amyloid beta protein ATM protein ATR protein checkpoint kinase 1 checkpoint kinase 2 cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase 4 cyclin dependent kinase 6 cyclin dependent kinase inhibitor 2D phosphate p 32 serine threonine amyloid beta protein CDK2 protein, human cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase inhibitor 2D DNA article cell nucleus cell survival conformational transition controlled study DNA damage DNA repair point mutation protein phosphorylation signal transduction ultraviolet radiation cell cycle cell line cell strain HEK293 DNA damage DNA repair drug effect gene expression regulation genetics human metabolism mutation phosphorylation protein transport radiation exposure Amyloid beta-Peptides Cell Cycle Cell Line Cell Survival Cisplatin Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinase Inhibitor p19 DNA DNA Damage DNA Repair Gene Expression Regulation HEK293 Cells Humans Mutation Phosphorylation Protein Transport Signal Transduction Ultraviolet Rays Marazita, Mariela C. Ogara, Maria Florencia Sonzogni, Silvina Verónica Martí, Marcelo Adrián Dusetti, Nelson J. Pignataro, Omar Pedro Cánepa, Eduardo Tomás CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
topic_facet	amyloid beta protein ATM protein ATR protein checkpoint kinase 1 checkpoint kinase 2 cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase 4 cyclin dependent kinase 6 cyclin dependent kinase inhibitor 2D phosphate p 32 serine threonine amyloid beta protein CDK2 protein, human cisplatin cyclic AMP dependent protein kinase cyclin dependent kinase 2 cyclin dependent kinase inhibitor 2D DNA article cell nucleus cell survival conformational transition controlled study DNA damage DNA repair point mutation protein phosphorylation signal transduction ultraviolet radiation cell cycle cell line cell strain HEK293 DNA damage DNA repair drug effect gene expression regulation genetics human metabolism mutation phosphorylation protein transport radiation exposure Amyloid beta-Peptides Cell Cycle Cell Line Cell Survival Cisplatin Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinase Inhibitor p19 DNA DNA Damage DNA Repair Gene Expression Regulation HEK293 Cells Humans Mutation Phosphorylation Protein Transport Signal Transduction Ultraviolet Rays
description	DNA damage triggers a phosphorylation-based signaling cascade known as the DNA damage response. p19INK4d, a member of the INK4 family of CDK4/6 inhibitors, has been reported to participate in the DNA damage response promoting DNA repair and cell survival. Here, we provide mechanistic insight into the activation mechanism of p19INK4d linked to the response to DNA damage. Results showed that p19INK4d becomes phosphorylated following UV radiation, b-amyloid peptide and cisplatin treatments. ATM-Chk2/ATR-Chk1 signaling pathways were found to be differentially involved in p19INK4d phosphorylation depending on the type of DNA damage. Two sequential phosphorylation events at serine 76 and threonine 141 were identified using p19INK4d single-point mutants in metabolic labeling assays with 32P-orthophosphate. CDK2 and PKA were found to participate in p19INK4d phosphorylation process and that they would mediate serine 76 and threonine 141 modifications respectively. Nuclear translocation of p19INK4d induced by DNA damage was shown to be dependent on serine 76 phosphorylation. Most importantly, both phosphorylation sites were found to be crucial for p19INK4d function in DNA repair and cell survival. In contrast, serine 76 and threonine 141 were dispensable for CDK4/6 inhibition highlighting the independence of p19INK4d functions, in agreement with our previous findings. These results constitute the first description of the activation mechanism of p19INK4d in response to genotoxic stress and demonstrate the functional relevance of this activation following DNA damage. © 2012 Marazita et al.
author	Marazita, Mariela C. Ogara, Maria Florencia Sonzogni, Silvina Verónica Martí, Marcelo Adrián Dusetti, Nelson J. Pignataro, Omar Pedro Cánepa, Eduardo Tomás
author_facet	Marazita, Mariela C. Ogara, Maria Florencia Sonzogni, Silvina Verónica Martí, Marcelo Adrián Dusetti, Nelson J. Pignataro, Omar Pedro Cánepa, Eduardo Tomás
author_sort	Marazita, Mariela C.
title	CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
title_short	CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
title_full	CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
title_fullStr	CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
title_full_unstemmed	CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response
title_sort	cdk2 and pka mediated-sequential phosphorylation is critical for p19ink4d function in the dna damage response
publishDate	2012
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v7_n4_p_Marazita http://hdl.handle.net/20.500.12110/paper_19326203_v7_n4_p_Marazita
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CDK2 and PKA mediated-sequential phosphorylation is critical for p19INK4d function in the DNA damage response

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