Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress

Jason L Parsons; Svetlana V Khoronenkova; Irina I Dianova; Nicola Ternette; Benedikt M Kessler; Pran K Datta; Grigory L Dianov

doi:10.1093/nar/gks909

Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress

Jason L Parsons, Svetlana V Khoronenkova, Irina I Dianova, Nicola Ternette, Benedikt M Kessler, Pran K Datta, Grigory L Dianov

Cancer and Genomic Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

We examined the mechanism regulating the cellular levels of PNKP, the major kinase/phosphatase involved in the repair of oxidative DNA damage, and find that it is controlled by ATM phosphorylation and ubiquitylation-dependent proteasomal degradation. We discovered that ATM-dependent phosphorylation of PNKP at serines 114 and 126 in response to oxidative DNA damage inhibits ubiquitylation-dependent proteasomal degradation of PNKP, and consequently increases PNKP stability that is required for DNA repair. We have also purified a novel Cul4A-DDB1 ubiquitin ligase complex responsible for PNKP ubiquitylation and identify serine-threonine kinase receptor associated protein (STRAP) as the adaptor protein that provides specificity of the complex to PNKP. Strap(-/-) mouse embryonic fibroblasts subsequently contain elevated cellular levels of PNKP, and show elevated resistance to oxidative DNA damage. These data demonstrate an important role for ATM and the Cul4A-DDB1-STRAP ubiquitin ligase in the regulation of the cellular levels of PNKP, and consequently in the repair of oxidative DNA damage.

Original language	English
Pages (from-to)	11404-15
Number of pages	12
Journal	Nucleic Acids Research
Volume	40
Issue number	22
DOIs	https://doi.org/10.1093/nar/gks909
Publication status	Published - Dec 2012

Keywords

Animals
Ataxia Telangiectasia Mutated Proteins
Carrier Proteins/metabolism
Cell Cycle Proteins/metabolism
Cullin Proteins/metabolism
DNA Damage
DNA Repair Enzymes/chemistry
DNA-Binding Proteins/metabolism
Enzyme Stability
HeLa Cells
Humans
Mice
Oxidative Stress
Phosphorylation
Phosphotransferases (Alcohol Group Acceptor)/chemistry
Proteasome Endopeptidase Complex/metabolism
Protein Serine-Threonine Kinases/metabolism
Tumor Suppressor Proteins/metabolism
Ubiquitin-Protein Ligases/isolation & purification
Ubiquitination

Access to Document

10.1093/nar/gks909

Cite this

@article{54dddfe932c14920957496e547434a0b,

title = "Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress",

abstract = "We examined the mechanism regulating the cellular levels of PNKP, the major kinase/phosphatase involved in the repair of oxidative DNA damage, and find that it is controlled by ATM phosphorylation and ubiquitylation-dependent proteasomal degradation. We discovered that ATM-dependent phosphorylation of PNKP at serines 114 and 126 in response to oxidative DNA damage inhibits ubiquitylation-dependent proteasomal degradation of PNKP, and consequently increases PNKP stability that is required for DNA repair. We have also purified a novel Cul4A-DDB1 ubiquitin ligase complex responsible for PNKP ubiquitylation and identify serine-threonine kinase receptor associated protein (STRAP) as the adaptor protein that provides specificity of the complex to PNKP. Strap(-/-) mouse embryonic fibroblasts subsequently contain elevated cellular levels of PNKP, and show elevated resistance to oxidative DNA damage. These data demonstrate an important role for ATM and the Cul4A-DDB1-STRAP ubiquitin ligase in the regulation of the cellular levels of PNKP, and consequently in the repair of oxidative DNA damage.",

keywords = "Animals, Ataxia Telangiectasia Mutated Proteins, Carrier Proteins/metabolism, Cell Cycle Proteins/metabolism, Cullin Proteins/metabolism, DNA Damage, DNA Repair Enzymes/chemistry, DNA-Binding Proteins/metabolism, Enzyme Stability, HeLa Cells, Humans, Mice, Oxidative Stress, Phosphorylation, Phosphotransferases (Alcohol Group Acceptor)/chemistry, Proteasome Endopeptidase Complex/metabolism, Protein Serine-Threonine Kinases/metabolism, Tumor Suppressor Proteins/metabolism, Ubiquitin-Protein Ligases/isolation & purification, Ubiquitination",

author = "Parsons, {Jason L} and Khoronenkova, {Svetlana V} and Dianova, {Irina I} and Nicola Ternette and Kessler, {Benedikt M} and Datta, {Pran K} and Dianov, {Grigory L}",

year = "2012",

month = dec,

doi = "10.1093/nar/gks909",

language = "English",

volume = "40",

pages = "11404--15",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "22",

}

TY - JOUR

T1 - Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress

AU - Parsons, Jason L

AU - Khoronenkova, Svetlana V

AU - Dianova, Irina I

AU - Ternette, Nicola

AU - Kessler, Benedikt M

AU - Datta, Pran K

AU - Dianov, Grigory L

PY - 2012/12

Y1 - 2012/12

N2 - We examined the mechanism regulating the cellular levels of PNKP, the major kinase/phosphatase involved in the repair of oxidative DNA damage, and find that it is controlled by ATM phosphorylation and ubiquitylation-dependent proteasomal degradation. We discovered that ATM-dependent phosphorylation of PNKP at serines 114 and 126 in response to oxidative DNA damage inhibits ubiquitylation-dependent proteasomal degradation of PNKP, and consequently increases PNKP stability that is required for DNA repair. We have also purified a novel Cul4A-DDB1 ubiquitin ligase complex responsible for PNKP ubiquitylation and identify serine-threonine kinase receptor associated protein (STRAP) as the adaptor protein that provides specificity of the complex to PNKP. Strap(-/-) mouse embryonic fibroblasts subsequently contain elevated cellular levels of PNKP, and show elevated resistance to oxidative DNA damage. These data demonstrate an important role for ATM and the Cul4A-DDB1-STRAP ubiquitin ligase in the regulation of the cellular levels of PNKP, and consequently in the repair of oxidative DNA damage.

AB - We examined the mechanism regulating the cellular levels of PNKP, the major kinase/phosphatase involved in the repair of oxidative DNA damage, and find that it is controlled by ATM phosphorylation and ubiquitylation-dependent proteasomal degradation. We discovered that ATM-dependent phosphorylation of PNKP at serines 114 and 126 in response to oxidative DNA damage inhibits ubiquitylation-dependent proteasomal degradation of PNKP, and consequently increases PNKP stability that is required for DNA repair. We have also purified a novel Cul4A-DDB1 ubiquitin ligase complex responsible for PNKP ubiquitylation and identify serine-threonine kinase receptor associated protein (STRAP) as the adaptor protein that provides specificity of the complex to PNKP. Strap(-/-) mouse embryonic fibroblasts subsequently contain elevated cellular levels of PNKP, and show elevated resistance to oxidative DNA damage. These data demonstrate an important role for ATM and the Cul4A-DDB1-STRAP ubiquitin ligase in the regulation of the cellular levels of PNKP, and consequently in the repair of oxidative DNA damage.

KW - Animals

KW - Ataxia Telangiectasia Mutated Proteins

KW - Carrier Proteins/metabolism

KW - Cell Cycle Proteins/metabolism

KW - Cullin Proteins/metabolism

KW - DNA Damage

KW - DNA Repair Enzymes/chemistry

KW - DNA-Binding Proteins/metabolism

KW - Enzyme Stability

KW - HeLa Cells

KW - Humans

KW - Mice

KW - Oxidative Stress

KW - Phosphorylation

KW - Phosphotransferases (Alcohol Group Acceptor)/chemistry

KW - Proteasome Endopeptidase Complex/metabolism

KW - Protein Serine-Threonine Kinases/metabolism

KW - Tumor Suppressor Proteins/metabolism

KW - Ubiquitin-Protein Ligases/isolation & purification

KW - Ubiquitination

U2 - 10.1093/nar/gks909

DO - 10.1093/nar/gks909

M3 - Article

C2 - 23042680

SN - 0305-1048

VL - 40

SP - 11404

EP - 11415

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 22

ER -

Phosphorylation of PNKP by ATM prevents its proteasomal degradation and enhances resistance to oxidative stress

Abstract

Keywords

Access to Document

Fingerprint

Cite this