ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability

Min Hwa Shin; Ming Yuan; Hao Zhang; Joseph B. Margolick; Mihoko Kai

doi:10.4161/cc.20161

ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability

Min Hwa Shin, Ming Yuan, Hao Zhang, Joseph B. Margolick, Mihoko Kai

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

11 Scopus citations

Abstract

Upon genotoxic stress and during normal S phase, ATM phosphorylates the checkpoint clamp protein Rad9 in a manner that depends on Ser272. Ser272 is the only known ATM-dependent phosphorylation site in human Rad9. However, Ser272 phosphorylation is not required for survival or checkpoint activation after DNA damage. The physiological function of Ser272 remains elusive. Here, we show that ATM-dependent Rad9^Ser272 phosphorylation requires the MRN complex and controls repair pathways. Furthermore, the mutant cells accumulate large numbers of chromosome breaks and induce gross chromosomal rearrangements. Our findings establish a new and unexpected role for ATM: it phosphorylates the checkpoint clamp in order to control repair pathways, thereby maintaining genomic integrity during unperturbed cell cycle and upon DNA damage.

Original language	English (US)
Pages (from-to)	1796-1803
Number of pages	8
Journal	Cell Cycle
Volume	11
Issue number	9
DOIs	https://doi.org/10.4161/cc.20161
State	Published - May 1 2012

Keywords

ATM
Checkpoint
Phosphorylation
Rad9
Repair

ASJC Scopus subject areas

Molecular Biology
Developmental Biology
Cell Biology

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10.4161/cc.20161

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title = "ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability",

abstract = "Upon genotoxic stress and during normal S phase, ATM phosphorylates the checkpoint clamp protein Rad9 in a manner that depends on Ser272. Ser272 is the only known ATM-dependent phosphorylation site in human Rad9. However, Ser272 phosphorylation is not required for survival or checkpoint activation after DNA damage. The physiological function of Ser272 remains elusive. Here, we show that ATM-dependent Rad9Ser272 phosphorylation requires the MRN complex and controls repair pathways. Furthermore, the mutant cells accumulate large numbers of chromosome breaks and induce gross chromosomal rearrangements. Our findings establish a new and unexpected role for ATM: it phosphorylates the checkpoint clamp in order to control repair pathways, thereby maintaining genomic integrity during unperturbed cell cycle and upon DNA damage.",

keywords = "ATM, Checkpoint, Phosphorylation, Rad9, Repair",

author = "Shin, {Min Hwa} and Ming Yuan and Hao Zhang and Margolick, {Joseph B.} and Mihoko Kai",

note = "Funding Information: GFP-based recombination assay. I-SceI expression plasmids We thank M. Jasin for providing us the DR293 cell line and (100 μg) were electroporated (250 V, 950 μF) into DR293 the I-SceI expression plasmid. This work was supported by a cells (5 x 106 cells).32 After electroporation, cells were plated in K01 award (5K01CA114027) from National Institute of Health non-selective media for 72 h and were recovered and concen-(NIH, M.K.). trated by centrifugation and resuspended in Opti-MEM media (Invitrogen) prior to flow cytometry. Cells were analyzed in FACSCalibur (BD Biosciences).",

year = "2012",

month = may,

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doi = "10.4161/cc.20161",

language = "English (US)",

volume = "11",

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journal = "Cell Cycle",

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T1 - ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability

AU - Shin, Min Hwa

AU - Yuan, Ming

AU - Zhang, Hao

AU - Margolick, Joseph B.

AU - Kai, Mihoko

N1 - Funding Information: GFP-based recombination assay. I-SceI expression plasmids We thank M. Jasin for providing us the DR293 cell line and (100 μg) were electroporated (250 V, 950 μF) into DR293 the I-SceI expression plasmid. This work was supported by a cells (5 x 106 cells).32 After electroporation, cells were plated in K01 award (5K01CA114027) from National Institute of Health non-selective media for 72 h and were recovered and concen-(NIH, M.K.). trated by centrifugation and resuspended in Opti-MEM media (Invitrogen) prior to flow cytometry. Cells were analyzed in FACSCalibur (BD Biosciences).

PY - 2012/5/1

Y1 - 2012/5/1

N2 - Upon genotoxic stress and during normal S phase, ATM phosphorylates the checkpoint clamp protein Rad9 in a manner that depends on Ser272. Ser272 is the only known ATM-dependent phosphorylation site in human Rad9. However, Ser272 phosphorylation is not required for survival or checkpoint activation after DNA damage. The physiological function of Ser272 remains elusive. Here, we show that ATM-dependent Rad9Ser272 phosphorylation requires the MRN complex and controls repair pathways. Furthermore, the mutant cells accumulate large numbers of chromosome breaks and induce gross chromosomal rearrangements. Our findings establish a new and unexpected role for ATM: it phosphorylates the checkpoint clamp in order to control repair pathways, thereby maintaining genomic integrity during unperturbed cell cycle and upon DNA damage.

AB - Upon genotoxic stress and during normal S phase, ATM phosphorylates the checkpoint clamp protein Rad9 in a manner that depends on Ser272. Ser272 is the only known ATM-dependent phosphorylation site in human Rad9. However, Ser272 phosphorylation is not required for survival or checkpoint activation after DNA damage. The physiological function of Ser272 remains elusive. Here, we show that ATM-dependent Rad9Ser272 phosphorylation requires the MRN complex and controls repair pathways. Furthermore, the mutant cells accumulate large numbers of chromosome breaks and induce gross chromosomal rearrangements. Our findings establish a new and unexpected role for ATM: it phosphorylates the checkpoint clamp in order to control repair pathways, thereby maintaining genomic integrity during unperturbed cell cycle and upon DNA damage.

KW - ATM

KW - Checkpoint

KW - Phosphorylation

KW - Rad9

KW - Repair

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ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability

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Keywords

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