Distinct roles of ATR and DNA-PKcs in triggering DNA damage responses in ATM-deficient cells

Nozomi Tomimatsu, Bipasha Mukherjee, Sandeep Burma

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

The cellular response to DNA double-strand breaks involves direct activation of ataxia telangiectasia mutated (ATM) and indirect activation of ataxia telangiectasia and Rad3 related (ATR) in an ATM/ Mre11/cell-cycle-dependent manner. Here, we report that the crucial checkpoint signalling proteins-p53, structural maintainance of chromosomes 1 (SMC1), p53 binding protein 1 (53BP1), checkpoint kinase (Chk)1 and Chk2-are phosphorylated rapidly by ATR in an ATM/Mre11/ cell-cycle-independent manner, albeit at low levels. We observed the sequential recruitment of replication protein A (RPA) and ATR to the sites of DNA damage in ATM-deficient cells, which provides a mechanistic basis for the observed phosphorylations. The recruitment of ATR and consequent phosphorylations do not require Mre11 but are dependent on Exo1. We show that these low levels of phosphorylation are biologically important, as ATM-deficient cells enforce an early G2/M checkpoint that is ATR-dependent. ATR is also essential for the late G2 accumulation that is peculiar to irradiated ATM-deficient cells. Interestingly, phosphorylation of KRAB associated protein 1 (KAP-1), a protein involved in chromatin remodelling, is mediated by DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in a spatio-temporal manner in addition to ATM. We posit that ATM substrates involved in cell-cycle checkpoint signalling can be minimally phosphorylated independently by ATR, while a small subset of proteins involved in chromatin remodelling are phosphorylated by DNA-PKcs in addition to ATM.

Original languageEnglish (US)
Pages (from-to)629-635
Number of pages7
JournalEMBO Reports
Volume10
Issue number6
DOIs
StatePublished - 2009

Fingerprint

DNA-Activated Protein Kinase
Ataxia Telangiectasia
Phosphorylation
DNA Damage
Catalytic Domain
DNA
Cells
Chromatin
Proteins
Chemical activation
Replication Protein A
Chromosomes
Carrier Proteins
Phosphotransferases
Chromatin Assembly and Disassembly
Substrates
Cell Cycle
Double-Stranded DNA Breaks

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Biochemistry

Cite this

Distinct roles of ATR and DNA-PKcs in triggering DNA damage responses in ATM-deficient cells. / Tomimatsu, Nozomi; Mukherjee, Bipasha; Burma, Sandeep.

In: EMBO Reports, Vol. 10, No. 6, 2009, p. 629-635.

Research output: Contribution to journalArticle

Tomimatsu, Nozomi ; Mukherjee, Bipasha ; Burma, Sandeep. / Distinct roles of ATR and DNA-PKcs in triggering DNA damage responses in ATM-deficient cells. In: EMBO Reports. 2009 ; Vol. 10, No. 6. pp. 629-635.
@article{d6f95def388f452ebc9695c71bdceedb,
title = "Distinct roles of ATR and DNA-PKcs in triggering DNA damage responses in ATM-deficient cells",
abstract = "The cellular response to DNA double-strand breaks involves direct activation of ataxia telangiectasia mutated (ATM) and indirect activation of ataxia telangiectasia and Rad3 related (ATR) in an ATM/ Mre11/cell-cycle-dependent manner. Here, we report that the crucial checkpoint signalling proteins-p53, structural maintainance of chromosomes 1 (SMC1), p53 binding protein 1 (53BP1), checkpoint kinase (Chk)1 and Chk2-are phosphorylated rapidly by ATR in an ATM/Mre11/ cell-cycle-independent manner, albeit at low levels. We observed the sequential recruitment of replication protein A (RPA) and ATR to the sites of DNA damage in ATM-deficient cells, which provides a mechanistic basis for the observed phosphorylations. The recruitment of ATR and consequent phosphorylations do not require Mre11 but are dependent on Exo1. We show that these low levels of phosphorylation are biologically important, as ATM-deficient cells enforce an early G2/M checkpoint that is ATR-dependent. ATR is also essential for the late G2 accumulation that is peculiar to irradiated ATM-deficient cells. Interestingly, phosphorylation of KRAB associated protein 1 (KAP-1), a protein involved in chromatin remodelling, is mediated by DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in a spatio-temporal manner in addition to ATM. We posit that ATM substrates involved in cell-cycle checkpoint signalling can be minimally phosphorylated independently by ATR, while a small subset of proteins involved in chromatin remodelling are phosphorylated by DNA-PKcs in addition to ATM.",
author = "Nozomi Tomimatsu and Bipasha Mukherjee and Sandeep Burma",
year = "2009",
doi = "10.1038/embor.2009.60",
language = "English (US)",
volume = "10",
pages = "629--635",
journal = "EMBO Reports",
issn = "1469-221X",
publisher = "Nature Publishing Group",
number = "6",

}

TY - JOUR

T1 - Distinct roles of ATR and DNA-PKcs in triggering DNA damage responses in ATM-deficient cells

AU - Tomimatsu, Nozomi

AU - Mukherjee, Bipasha

AU - Burma, Sandeep

PY - 2009

Y1 - 2009

N2 - The cellular response to DNA double-strand breaks involves direct activation of ataxia telangiectasia mutated (ATM) and indirect activation of ataxia telangiectasia and Rad3 related (ATR) in an ATM/ Mre11/cell-cycle-dependent manner. Here, we report that the crucial checkpoint signalling proteins-p53, structural maintainance of chromosomes 1 (SMC1), p53 binding protein 1 (53BP1), checkpoint kinase (Chk)1 and Chk2-are phosphorylated rapidly by ATR in an ATM/Mre11/ cell-cycle-independent manner, albeit at low levels. We observed the sequential recruitment of replication protein A (RPA) and ATR to the sites of DNA damage in ATM-deficient cells, which provides a mechanistic basis for the observed phosphorylations. The recruitment of ATR and consequent phosphorylations do not require Mre11 but are dependent on Exo1. We show that these low levels of phosphorylation are biologically important, as ATM-deficient cells enforce an early G2/M checkpoint that is ATR-dependent. ATR is also essential for the late G2 accumulation that is peculiar to irradiated ATM-deficient cells. Interestingly, phosphorylation of KRAB associated protein 1 (KAP-1), a protein involved in chromatin remodelling, is mediated by DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in a spatio-temporal manner in addition to ATM. We posit that ATM substrates involved in cell-cycle checkpoint signalling can be minimally phosphorylated independently by ATR, while a small subset of proteins involved in chromatin remodelling are phosphorylated by DNA-PKcs in addition to ATM.

AB - The cellular response to DNA double-strand breaks involves direct activation of ataxia telangiectasia mutated (ATM) and indirect activation of ataxia telangiectasia and Rad3 related (ATR) in an ATM/ Mre11/cell-cycle-dependent manner. Here, we report that the crucial checkpoint signalling proteins-p53, structural maintainance of chromosomes 1 (SMC1), p53 binding protein 1 (53BP1), checkpoint kinase (Chk)1 and Chk2-are phosphorylated rapidly by ATR in an ATM/Mre11/ cell-cycle-independent manner, albeit at low levels. We observed the sequential recruitment of replication protein A (RPA) and ATR to the sites of DNA damage in ATM-deficient cells, which provides a mechanistic basis for the observed phosphorylations. The recruitment of ATR and consequent phosphorylations do not require Mre11 but are dependent on Exo1. We show that these low levels of phosphorylation are biologically important, as ATM-deficient cells enforce an early G2/M checkpoint that is ATR-dependent. ATR is also essential for the late G2 accumulation that is peculiar to irradiated ATM-deficient cells. Interestingly, phosphorylation of KRAB associated protein 1 (KAP-1), a protein involved in chromatin remodelling, is mediated by DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in a spatio-temporal manner in addition to ATM. We posit that ATM substrates involved in cell-cycle checkpoint signalling can be minimally phosphorylated independently by ATR, while a small subset of proteins involved in chromatin remodelling are phosphorylated by DNA-PKcs in addition to ATM.

UR - http://www.scopus.com/inward/record.url?scp=67349288164&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67349288164&partnerID=8YFLogxK

U2 - 10.1038/embor.2009.60

DO - 10.1038/embor.2009.60

M3 - Article

C2 - 19444312

AN - SCOPUS:67349288164

VL - 10

SP - 629

EP - 635

JO - EMBO Reports

JF - EMBO Reports

SN - 1469-221X

IS - 6

ER -