460dab82d8618dc20e36fb8e20cf6cea pmc_33076429.pdf cd1bfa077ba58bbca85676cc2360013d224f8409 pmc_33076429.pdf 27a80ba0f82329b4e12b2c18ec834be3f50d74638f4afadbb2c9a71391a06852 pmc_33076429.pdf Title: VRK1 Phosphorylates Tip60/KAT5 and Is Required for H4K16 Acetylation in Response to DNA Damage Subject: Dynamic remodeling of chromatin requires acetylation and methylation of histones, frequently affecting the same lysine residue. These alternative epigenetic modifications require the coordination of enzymes, writers and erasers, mediating them such as acetylases and deacetylases. In cells in G0/G1, DNA damage induced by doxorubicin causes an increase in histone H4K16ac, a marker of chromatin relaxation. In this context, we studied the role that VRK1, a chromatin kinase activated by DNA damage, plays in this early step. VRK1 depletion or MG149, a Tip60/KAT5 inhibitor, cause a loss of H4K16ac. DNA damage induces the phosphorylation of Tip60 mediated by VRK1 in the chromatin fraction. VRK1 directly interacts with and phosphorylates Tip60. Furthermore, the phosphorylation of Tip60 induced by doxorubicin is lost by depletion of VRK1 in both ATM +/+ and ATM-/- cells. Kinase-active VRK1, but not kinase-dead VRK1, rescues Tip60 phosphorylation induced by DNA damage independently of ATM. The Tip60 phosphorylation by VRK1 is necessary for the activating acetylation of ATM, and subsequent ATM autophosphorylation, and both are lost by VRK1 depletion. These results support that the VRK1 chromatin kinase is an upstream regulator of the initial acetylation of histones, and an early step in DNA damage responses (DDR). Keywords: phosphorylation; histone H4; acetylation; DNA-damage response; nucleosomal histone kinase-1 Author: Raúl García-González, Patricia Morejón-García, Ignacio Campillo-Marcos, Marcella Salzano and Pedro A. 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no 928 0 Arial CID TrueType Identity-H yes no yes 931 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 985 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 989 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 993 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 997 0 Times New Roman TrueType WinAnsi no no no 1004 0 ABCDEE+Calibri,Bold TrueType WinAnsi yes yes no 1009 0 Arial,Bold TrueType WinAnsi no no no 1013 0 VSHNAE+Rpcxr Type 1 Builtin yes yes no 1041 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-06-24 02:33:17 CEST RepresentationInformation: pmc_33076429.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2020-11-23 03:38:02 CET Size: 7252576 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1405 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: DNA Damage Induces Phosphorylation of Tip60 and Acetylation of H4K16 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Kinase-Active VRK1 Rescues H4K16 Acetylation Induced by DNA Damage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: VRK1 Directly Interacts with and Phosphorylates Tip60 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Kinase-Active VRK1 Mediates the Phosphorylation of Tip60 Induced by DNA-Damage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: VRK1 Depletion Prevents the Accumulation in Chromatin of Phospho-Tip60 Induced by DNA Damage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: The Phosphorylation of Tip60 by VRK1 is Induced by Doxorubicin in ATM Null Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: ATM Acetylation by Tip60 Is Insensitive to ATM Kinase Inhibitors but Is Lost by VRK1 Depletion or Tip60 Inhibition Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: The Kinase Activity of VRK1 Is Necessary to Rescue ATM Acetylation and Phosphorylation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Children: Item: Title: Reagents Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Plasmids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Cell Lines, Culture and Transfections Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: VRK1 Depletion by siRNA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Antibodies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Immunoblots Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Immunoprecipitations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: In Vitro VRK1-Tip60 Protein Interaction and Kinase Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Immunofluorescence and Confocal Microscopy Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Nucleoplasm-Chromatin Fractionation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: TUNEL Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Database Submission Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Info: Title: VRK1 Phosphorylates Tip60/KAT5 and Is Required for H4K16 Acetylation in Response to DNA Damage Author: Raúl García-González, Patricia Morejón-García, Ignacio Campillo-Marcos, Marcella Salzano and Pedro A. Lazo Subject: Dynamic remodeling of chromatin requires acetylation and methylation of histones, frequently affecting the same lysine residue. These alternative epigenetic modifications require the coordination of enzymes, writers and erasers, mediating them such as acetylases and deacetylases. In cells in G0/G1, DNA damage induced by doxorubicin causes an increase in histone H4K16ac, a marker of chromatin relaxation. In this context, we studied the role that VRK1, a chromatin kinase activated by DNA damage, plays in this early step. VRK1 depletion or MG149, a Tip60/KAT5 inhibitor, cause a loss of H4K16ac. DNA damage induces the phosphorylation of Tip60 mediated by VRK1 in the chromatin fraction. VRK1 directly interacts with and phosphorylates Tip60. Furthermore, the phosphorylation of Tip60 induced by doxorubicin is lost by depletion of VRK1 in both ATM +/+ and ATM-/- cells. Kinase-active VRK1, but not kinase-dead VRK1, rescues Tip60 phosphorylation induced by DNA damage independently of ATM. The Tip60 phosphorylation by VRK1 is necessary for the activating acetylation of ATM, and subsequent ATM autophosphorylation, and both are lost by VRK1 depletion. These results support that the VRK1 chromatin kinase is an upstream regulator of the initial acetylation of histones, and an early step in DNA damage responses (DDR). 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325 Checksum: 3797320f Type: CRC32 Checksum: 460dab82d8618dc20e36fb8e20cf6cea Type: MD5 Checksum: cd1bfa077ba58bbca85676cc2360013d224f8409 Type: SHA-1