Early Chk1 Phosphorylation Is Driven by Temozolomide-Induced, DNA Double Strand Break- and Mismatch Repair-Independent DNA Damage

Motokazu Ito, Shigeo Oba, Karin Gaensler, Sabrina M. Ronen, Joydeep Mukherjee, Russell O. Pieper

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Temozolomide (TMZ) is a DNA methylating agent used to treat brain cancer. TMZ-induced O6-methylguanine adducts, in the absence of repair by O6-methylguanine DNA methyltransferase (MGMT), mispair during DNA replication and trigger cycles of futile mismatch repair (MMR). Futile MMR in turn leads to the formation of DNA single and double strand breaks, Chk1 and Chk2 phosphorylation/activation, cell cycle arrest, and ultimately cell death. Although both pChk1 and pChk2 are considered to be biomarkers of TMZ-induced DNA damage, cell-cycle arrest, and TMZ induced cytotoxicity, we found that levels of pChk1 (ser345), its downstream target pCdc25C (ser216), and the activity of its upstream activator ATR, were elevated within 3 hours of TMZ exposure, long before the onset of TMZ-induced DNA double strand breaks, Chk2 phosphorylation/activation, and cell cycle arrest. Furthermore, TMZ-induced early phosphorylation of Chk1 was noted in glioma cells regardless of whether they were MGMT-proficient or MGMT-deficient, and regardless of their MMR status. Early Chk1 phosphorylation was not associated with TMZ-induced reactive oxygen species, but was temporally associated with TMZ-induced alkalai-labile DNA damage produced by the non-O6-methylguanine DNA adducts and which, like Chk1 phosphorylation, was transient in MGMT-proficient cells but persistent in MGMT-deficient cells. These results re-define the TMZ-induced DNA damage response, and show that Chk1 phosphorylation is driven by TMZ-induced mismatch repair-independent DNA damage independently of DNA double strand breaks, Chk2 activation, and cell cycle arrest, and as such is a suboptimal biomarker of TMZ-induced drug action.

Original languageEnglish
Article numbere62351
JournalPloS one
Volume8
Issue number5
DOIs
Publication statusPublished - 07-05-2013

Fingerprint

temozolomide
Phosphorylation
DNA Mismatch Repair
Double-Stranded DNA Breaks
DNA damage
DNA Damage
phosphorylation
Repair
methyltransferases
DNA
Methyltransferases
Cell Cycle Checkpoints
Cells
biomarkers
Chemical activation
Biomarkers
DNA adducts
Substrate Cycling
DNA replication
cells

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Ito, Motokazu ; Oba, Shigeo ; Gaensler, Karin ; Ronen, Sabrina M. ; Mukherjee, Joydeep ; Pieper, Russell O. / Early Chk1 Phosphorylation Is Driven by Temozolomide-Induced, DNA Double Strand Break- and Mismatch Repair-Independent DNA Damage. In: PloS one. 2013 ; Vol. 8, No. 5.
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title = "Early Chk1 Phosphorylation Is Driven by Temozolomide-Induced, DNA Double Strand Break- and Mismatch Repair-Independent DNA Damage",
abstract = "Temozolomide (TMZ) is a DNA methylating agent used to treat brain cancer. TMZ-induced O6-methylguanine adducts, in the absence of repair by O6-methylguanine DNA methyltransferase (MGMT), mispair during DNA replication and trigger cycles of futile mismatch repair (MMR). Futile MMR in turn leads to the formation of DNA single and double strand breaks, Chk1 and Chk2 phosphorylation/activation, cell cycle arrest, and ultimately cell death. Although both pChk1 and pChk2 are considered to be biomarkers of TMZ-induced DNA damage, cell-cycle arrest, and TMZ induced cytotoxicity, we found that levels of pChk1 (ser345), its downstream target pCdc25C (ser216), and the activity of its upstream activator ATR, were elevated within 3 hours of TMZ exposure, long before the onset of TMZ-induced DNA double strand breaks, Chk2 phosphorylation/activation, and cell cycle arrest. Furthermore, TMZ-induced early phosphorylation of Chk1 was noted in glioma cells regardless of whether they were MGMT-proficient or MGMT-deficient, and regardless of their MMR status. Early Chk1 phosphorylation was not associated with TMZ-induced reactive oxygen species, but was temporally associated with TMZ-induced alkalai-labile DNA damage produced by the non-O6-methylguanine DNA adducts and which, like Chk1 phosphorylation, was transient in MGMT-proficient cells but persistent in MGMT-deficient cells. These results re-define the TMZ-induced DNA damage response, and show that Chk1 phosphorylation is driven by TMZ-induced mismatch repair-independent DNA damage independently of DNA double strand breaks, Chk2 activation, and cell cycle arrest, and as such is a suboptimal biomarker of TMZ-induced drug action.",
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Early Chk1 Phosphorylation Is Driven by Temozolomide-Induced, DNA Double Strand Break- and Mismatch Repair-Independent DNA Damage. / Ito, Motokazu; Oba, Shigeo; Gaensler, Karin; Ronen, Sabrina M.; Mukherjee, Joydeep; Pieper, Russell O.

In: PloS one, Vol. 8, No. 5, e62351, 07.05.2013.

Research output: Contribution to journalArticle

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T1 - Early Chk1 Phosphorylation Is Driven by Temozolomide-Induced, DNA Double Strand Break- and Mismatch Repair-Independent DNA Damage

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