The Complexity of Phosphorylated H2AX Foci Formation and DNA Repair Assembly at DNA Double-strand Breaks

Overview

Journal Cell Cycle

Specialty Cell Biology

Date 2010 Jan 5

PMID 20046100

Citations 71

Authors

Asako J Nakamura

V Ashutosh Rao

Yves Pommier

William M Bonner

Affiliations

Soon will be listed here.

Abstract

The maintenance of genome stability requires efficient DNA double-stranded break (DSB) repair mediated by the phosphorylation of multiple histone H2AX molecules near the break sites. The phosphorylated H2AX (gammaH2AX) molecules form foci covering many megabases of chromatin. The formation of gamma-H2AX foci is critical for efficient DNA damage response (DDR) and for the maintenance of genome stability, however, the mechanisms of protein organization in foci is largely unknown. To investigate the nature of gammaH2AX foci formation, we analyzed the distribution of gammaH2AX and other DDR proteins at DSB sites using a variety of techniques to visualize, expand and partially disrupt chromatin. We report here that gammaH2AX foci change composition during the cell cycle, with proteins 53BP1, NBS1 and MRE11 dissociating from foci in G(2) and mitosis to return at the beginning of the following G(1). In contrast, MDC1 remained colocalized with gamma-H2AX during mitosis. In addition, while gammaH2AX was found to span large domains flanking DSB sites, 53BP1 and NBS1 were more localized and MDC1 colocalized in doublets in foci. H2AX and MDC1 were found to be involved in chromatin relaxation after DSB formation. Our data demonstrates that the DSB repair focus is a heterogeneous and dynamic structure containing internal complexity.

Citing Articles

Werner helicase interacting protein 1 contributes to G-quadruplex processing in human cells.

Hegedus L, Toth A, Harami G, Palinkas J, Karatayeva N, Sajben-Nagy E Sci Rep. 2024; 14(1):15740.

PMID: 38977862 PMC: 11231340. DOI: 10.1038/s41598-024-66425-y.

BUB1 regulates non-homologous end joining pathway to mediate radioresistance in triple-negative breast cancer.

Sriramulu S, Thoidingjam S, Chen W, Hassan O, Siddiqui F, Brown S J Exp Clin Cancer Res. 2024; 43(1):163.

PMID: 38863037 PMC: 11167950. DOI: 10.1186/s13046-024-03086-9.

Senescence-Associated Heterochromatin Foci Suppress γ-H2AX Focus Formation Induced by Radiation Exposure.

Oizumi T, Suzuki T, Kobayashi J, Nakamura A Int J Mol Sci. 2024; 25(6).

PMID: 38542327 PMC: 10969922. DOI: 10.3390/ijms25063355.

Current Strategies for Increasing Knock-In Efficiency in CRISPR/Cas9-Based Approaches.

Leal A, Herreno-Pachon A, Benincore-Florez E, Karunathilaka A, Tomatsu S Int J Mol Sci. 2024; 25(5).

PMID: 38473704 PMC: 10931195. DOI: 10.3390/ijms25052456.

Tumor targeted alpha particle therapy with an actinium-225 labelled antibody for carbonic anhydrase IX.

Morgan K, Wichmann C, Osellame L, Cao Z, Guo N, Scott A Chem Sci. 2024; 15(9):3372-3381.

PMID: 38425522 PMC: 10901495. DOI: 10.1039/d3sc06365h.

References

Zha S, Boboila C, Alt F . Mre11: roles in DNA repair beyond homologous recombination. Nat Struct Mol Biol. 2009; 16(8):798-800. DOI: 10.1038/nsmb0809-798. View

Takahashi K, Kaneko I . Changes in nuclease sensitivity of mammalian cells after irradiation with 60Co gamma-rays. Int J Radiat Biol Relat Stud Phys Chem Med. 1985; 48(3):389-95. DOI: 10.1080/09553008514551391. View

Di Masi A, Viganotti M, Polticelli F, Ascenzi P, Tanzarella C, Antoccia A . The R215W mutation in NBS1 impairs gamma-H2AX binding and affects DNA repair: molecular bases for the severe phenotype of 657del5/R215W Nijmegen breakage syndrome patients. Biochem Biophys Res Commun. 2008; 369(3):835-40. DOI: 10.1016/j.bbrc.2008.02.129. View

Carbone R, Pearson M, Minucci S, Pelicci P . PML NBs associate with the hMre11 complex and p53 at sites of irradiation induced DNA damage. Oncogene. 2002; 21(11):1633-40. DOI: 10.1038/sj.onc.1205227. View

Stucki M, Clapperton J, Mohammad D, Yaffe M, Smerdon S, Jackson S . MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell. 2005; 123(7):1213-26. DOI: 10.1016/j.cell.2005.09.038. View

Kolas N, Chapman J, Nakada S, Ylanko J, Chahwan R, Sweeney F . Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase. Science. 2007; 318(5856):1637-40. PMC: 2430610. DOI: 10.1126/science.1150034. View

Sullivan B, Karpen G . Centromeric chromatin exhibits a histone modification pattern that is distinct from both euchromatin and heterochromatin. Nat Struct Mol Biol. 2004; 11(11):1076-83. PMC: 1283111. DOI: 10.1038/nsmb845. View

Hoeijmakers J . Genome maintenance mechanisms for preventing cancer. Nature. 2001; 411(6835):366-74. DOI: 10.1038/35077232. View

Stiff T, Walker S, Cerosaletti K, Goodarzi A, Petermann E, Concannon P . ATR-dependent phosphorylation and activation of ATM in response to UV treatment or replication fork stalling. EMBO J. 2006; 25(24):5775-82. PMC: 1698893. DOI: 10.1038/sj.emboj.7601446. View

10.

Nelson G, Buhmann M, von Zglinicki T . DNA damage foci in mitosis are devoid of 53BP1. Cell Cycle. 2009; 8(20):3379-83. DOI: 10.4161/cc.8.20.9857. View

11.

Rogakou E, Boon C, Redon C, Bonner W . Megabase chromatin domains involved in DNA double-strand breaks in vivo. J Cell Biol. 1999; 146(5):905-16. PMC: 2169482. DOI: 10.1083/jcb.146.5.905. View

12.

Xie A, Hartlerode A, Stucki M, Odate S, Puget N, Kwok A . Distinct roles of chromatin-associated proteins MDC1 and 53BP1 in mammalian double-strand break repair. Mol Cell. 2007; 28(6):1045-57. PMC: 2275782. DOI: 10.1016/j.molcel.2007.12.005. View

13.

Kusch T, Florens L, MacDonald W, Swanson S, Glaser R, Yates 3rd J . Acetylation by Tip60 is required for selective histone variant exchange at DNA lesions. Science. 2004; 306(5704):2084-7. DOI: 10.1126/science.1103455. View

14.

Rao V, Fan A, Meng L, Doe C, North P, Hickson I . Phosphorylation of BLM, dissociation from topoisomerase IIIalpha, and colocalization with gamma-H2AX after topoisomerase I-induced replication damage. Mol Cell Biol. 2005; 25(20):8925-37. PMC: 1265790. DOI: 10.1128/MCB.25.20.8925-8937.2005. View

15.

Bonner W, Redon C, Dickey J, Nakamura A, Sedelnikova O, Solier S . GammaH2AX and cancer. Nat Rev Cancer. 2008; 8(12):957-67. PMC: 3094856. DOI: 10.1038/nrc2523. View

16.

Goodarzi A, Noon A, Deckbar D, Ziv Y, Shiloh Y, Lobrich M . ATM signaling facilitates repair of DNA double-strand breaks associated with heterochromatin. Mol Cell. 2008; 31(2):167-77. DOI: 10.1016/j.molcel.2008.05.017. View

17.

Morrison A, Highland J, Krogan N, Arbel-Eden A, Greenblatt J, Haber J . INO80 and gamma-H2AX interaction links ATP-dependent chromatin remodeling to DNA damage repair. Cell. 2004; 119(6):767-75. DOI: 10.1016/j.cell.2004.11.037. View

18.

Paull T, Rogakou E, Yamazaki V, Kirchgessner C, Gellert M, Bonner W . A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol. 2000; 10(15):886-95. DOI: 10.1016/s0960-9822(00)00610-2. View

19.

Berkovich E, Monnat Jr R, Kastan M . Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair. Nat Cell Biol. 2007; 9(6):683-90. DOI: 10.1038/ncb1599. View

20.

Dickey J, Redon C, Nakamura A, Baird B, Sedelnikova O, Bonner W . H2AX: functional roles and potential applications. Chromosoma. 2009; 118(6):683-92. PMC: 3094848. DOI: 10.1007/s00412-009-0234-4. View