A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery

Michael Christopher Keogh; Jung Ae Kim; Michael Downey; Jeffrey Fillingham; Dipanjan Chowdhury; Jacob C. Harrison; Megumi Onishi; Nira Datta; Sarah Galicia; Andrew Emili; Judy Lieberman; Xuetong Shen; Stephen Buratowski; James E. Haber; Daniel Durocher; Jack F. Greenblatt; Nevan J. Krogan

doi:10.1038/nature04384

A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery

Michael Christopher Keogh, Jung Ae Kim, Michael Downey, Jeffrey Fillingham, Dipanjan Chowdhury, Jacob C. Harrison, Megumi Onishi, Nira Datta, Sarah Galicia, Andrew Emili, Judy Lieberman, Xuetong Shen, Stephen Buratowski, James E. Haber, Daniel Durocher, Jack F. Greenblatt, Nevan J. Krogan

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407 Scopus citations

Abstract

One of the earliest marks of a double-strand break (DSB) in eukaryotes is serine phosphorylation of the histone variant H2AX at the carboxy-terminal SQE motif to create γH2AX-containing nucleosomes¹. Budding-yeast histone H2A is phosphorylated in a similar manner by the checkpoint kinases Tel1 and Mec1 (ref. 2; orthologous to mammalian ATM and ATR, respectively) over a 50-kilobase region surrounding the DSB³. This modification is important for recruiting numerous DSB-recognition and repair factors to the break site, including DNA damage checkpoint proteins^4,5, chromatin remodellers⁶ and cohesins^7,8. Multiple mechanisms for eliminating γH2AX as DNA repair completes are possible, including removal by histone exchange followed potentially by degradation, or, alternatively, dephosphorylation. Here we describe a three-protein complex (HTP-C, for histone H2A phosphatase complex) containing the phosphatase Pph3 that regulates the phosphorylation status of γH2AX in vivo and efficiently dephosphorylates γH2AX in vitro. γH2AX is lost from chromatin surrounding a DSB independently of the HTP-C, indicating that the phosphatase targets γH2AX after its displacement from DNA. The dephosphorylation of γH2AX by the HTP-C is necessary for efficient recovery from the DNA damage checkpoint.

Original language	English
Pages (from-to)	497-501
Number of pages	5
Journal	Nature
Volume	439
Issue number	7075
DOIs	https://doi.org/10.1038/nature04384
State	Published - 26 Jan 2006
Externally published	Yes

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Keogh, M. C., Kim, J. A., Downey, M., Fillingham, J., Chowdhury, D., Harrison, J. C., Onishi, M., Datta, N., Galicia, S., Emili, A., Lieberman, J., Shen, X., Buratowski, S., Haber, J. E., Durocher, D., Greenblatt, J. F., & Krogan, N. J. (2006). A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery. Nature, 439(7075), 497-501. https://doi.org/10.1038/nature04384

@article{1a138d08f6ea4cfea7195690dcbb9c43,

title = "A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery",

abstract = "One of the earliest marks of a double-strand break (DSB) in eukaryotes is serine phosphorylation of the histone variant H2AX at the carboxy-terminal SQE motif to create γH2AX-containing nucleosomes1. Budding-yeast histone H2A is phosphorylated in a similar manner by the checkpoint kinases Tel1 and Mec1 (ref. 2; orthologous to mammalian ATM and ATR, respectively) over a 50-kilobase region surrounding the DSB3. This modification is important for recruiting numerous DSB-recognition and repair factors to the break site, including DNA damage checkpoint proteins4,5, chromatin remodellers6 and cohesins7,8. Multiple mechanisms for eliminating γH2AX as DNA repair completes are possible, including removal by histone exchange followed potentially by degradation, or, alternatively, dephosphorylation. Here we describe a three-protein complex (HTP-C, for histone H2A phosphatase complex) containing the phosphatase Pph3 that regulates the phosphorylation status of γH2AX in vivo and efficiently dephosphorylates γH2AX in vitro. γH2AX is lost from chromatin surrounding a DSB independently of the HTP-C, indicating that the phosphatase targets γH2AX after its displacement from DNA. The dephosphorylation of γH2AX by the HTP-C is necessary for efficient recovery from the DNA damage checkpoint.",

author = "Keogh, {Michael Christopher} and Kim, {Jung Ae} and Michael Downey and Jeffrey Fillingham and Dipanjan Chowdhury and Harrison, {Jacob C.} and Megumi Onishi and Nira Datta and Sarah Galicia and Andrew Emili and Judy Lieberman and Xuetong Shen and Stephen Buratowski and Haber, {James E.} and Daniel Durocher and Greenblatt, {Jack F.} and Krogan, {Nevan J.}",

note = "Funding Information: Acknowledgements We thank C. J. Ingles for critically reading the manuscript and for discussion; C. Redon, W. Hofer and N. Lowndes for antibodies; E. O{\textquoteright}Shea and J. Weissman for yeast strains; and X. Wu, G. Zhong and X. Guo for technical assistance. N.J.K. was supported by a Doctoral Fellowship from the Canadian Institutes of Health Research (CIHR). J.C.H. and D.C. are fellows of the Leukemia and Lymphoma Society. D.D. is a recipient of the Hitchings-Elion Fellowship of the Burroughs-Wellcome Fund and a Canada Research Chair (tier II) in Proteomics, Bioinformatics and Functional Genomics. This research was supported by grants to J.F.G. from the CIHR and the Ontario Genomics Institute with funds from Genome Canada, to D.D. by the CIHR, to J.E.H. by the National Institutes of Health (NIH), and to S.B. by the NIH.",

year = "2006",

month = jan,

day = "26",

doi = "10.1038/nature04384",

language = "English",

volume = "439",

pages = "497--501",

journal = "Nature",

issn = "0028-0836",

publisher = "Nature Research",

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Keogh, MC, Kim, JA, Downey, M, Fillingham, J, Chowdhury, D, Harrison, JC, Onishi, M, Datta, N, Galicia, S, Emili, A, Lieberman, J, Shen, X, Buratowski, S, Haber, JE, Durocher, D, Greenblatt, JF & Krogan, NJ 2006, 'A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery', Nature, vol. 439, no. 7075, pp. 497-501. https://doi.org/10.1038/nature04384

TY - JOUR

T1 - A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery

AU - Keogh, Michael Christopher

AU - Kim, Jung Ae

AU - Downey, Michael

AU - Fillingham, Jeffrey

AU - Chowdhury, Dipanjan

AU - Harrison, Jacob C.

AU - Onishi, Megumi

AU - Datta, Nira

AU - Galicia, Sarah

AU - Emili, Andrew

AU - Lieberman, Judy

AU - Shen, Xuetong

AU - Buratowski, Stephen

AU - Haber, James E.

AU - Durocher, Daniel

AU - Greenblatt, Jack F.

AU - Krogan, Nevan J.

N1 - Funding Information: Acknowledgements We thank C. J. Ingles for critically reading the manuscript and for discussion; C. Redon, W. Hofer and N. Lowndes for antibodies; E. O’Shea and J. Weissman for yeast strains; and X. Wu, G. Zhong and X. Guo for technical assistance. N.J.K. was supported by a Doctoral Fellowship from the Canadian Institutes of Health Research (CIHR). J.C.H. and D.C. are fellows of the Leukemia and Lymphoma Society. D.D. is a recipient of the Hitchings-Elion Fellowship of the Burroughs-Wellcome Fund and a Canada Research Chair (tier II) in Proteomics, Bioinformatics and Functional Genomics. This research was supported by grants to J.F.G. from the CIHR and the Ontario Genomics Institute with funds from Genome Canada, to D.D. by the CIHR, to J.E.H. by the National Institutes of Health (NIH), and to S.B. by the NIH.

PY - 2006/1/26

Y1 - 2006/1/26

N2 - One of the earliest marks of a double-strand break (DSB) in eukaryotes is serine phosphorylation of the histone variant H2AX at the carboxy-terminal SQE motif to create γH2AX-containing nucleosomes1. Budding-yeast histone H2A is phosphorylated in a similar manner by the checkpoint kinases Tel1 and Mec1 (ref. 2; orthologous to mammalian ATM and ATR, respectively) over a 50-kilobase region surrounding the DSB3. This modification is important for recruiting numerous DSB-recognition and repair factors to the break site, including DNA damage checkpoint proteins4,5, chromatin remodellers6 and cohesins7,8. Multiple mechanisms for eliminating γH2AX as DNA repair completes are possible, including removal by histone exchange followed potentially by degradation, or, alternatively, dephosphorylation. Here we describe a three-protein complex (HTP-C, for histone H2A phosphatase complex) containing the phosphatase Pph3 that regulates the phosphorylation status of γH2AX in vivo and efficiently dephosphorylates γH2AX in vitro. γH2AX is lost from chromatin surrounding a DSB independently of the HTP-C, indicating that the phosphatase targets γH2AX after its displacement from DNA. The dephosphorylation of γH2AX by the HTP-C is necessary for efficient recovery from the DNA damage checkpoint.

AB - One of the earliest marks of a double-strand break (DSB) in eukaryotes is serine phosphorylation of the histone variant H2AX at the carboxy-terminal SQE motif to create γH2AX-containing nucleosomes1. Budding-yeast histone H2A is phosphorylated in a similar manner by the checkpoint kinases Tel1 and Mec1 (ref. 2; orthologous to mammalian ATM and ATR, respectively) over a 50-kilobase region surrounding the DSB3. This modification is important for recruiting numerous DSB-recognition and repair factors to the break site, including DNA damage checkpoint proteins4,5, chromatin remodellers6 and cohesins7,8. Multiple mechanisms for eliminating γH2AX as DNA repair completes are possible, including removal by histone exchange followed potentially by degradation, or, alternatively, dephosphorylation. Here we describe a three-protein complex (HTP-C, for histone H2A phosphatase complex) containing the phosphatase Pph3 that regulates the phosphorylation status of γH2AX in vivo and efficiently dephosphorylates γH2AX in vitro. γH2AX is lost from chromatin surrounding a DSB independently of the HTP-C, indicating that the phosphatase targets γH2AX after its displacement from DNA. The dephosphorylation of γH2AX by the HTP-C is necessary for efficient recovery from the DNA damage checkpoint.

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U2 - 10.1038/nature04384

DO - 10.1038/nature04384

M3 - Article

C2 - 16299494

AN - SCOPUS:31444445458

SN - 0028-0836

VL - 439

SP - 497

EP - 501

JO - Nature

JF - Nature

IS - 7075

ER -

A phosphatase complex that dephosphorylates γH2AX regulates DNA damage checkpoint recovery

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