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Title: A single double-strand break system reveals repair dynamics and mechanisms in heterochromatin and euchromatin

Journal Article · · Genes & Development
 [1];  [2];  [3];  [1];  [1];  [3];  [2];  [4];  [5]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Yale School of Medicine, New Haven, CT (United States)
  3. Netherlands Cancer Inst., Amsterdam (Netherlands)
  4. Georgetown Univ. Medical Center, Washington, DC (United States)
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)

Repair of DNA double-strand breaks (DSBs) must be properly orchestrated in diverse chromatin regions to maintain genome stability. The choice between two main DSB repair pathways, nonhomologous end-joining (NHEJ) and homologous recombination (HR), is regulated by the cell cycle as well as chromatin context.Pericentromeric heterochromatin forms a distinct nuclear domain that is enriched for repetitive DNA sequences that pose significant challenges for genome stability. Heterochromatic DSBs display specialized temporal and spatial dynamics that differ from euchromatic DSBs. Although HR is thought to be the main pathway used to repair heterochromatic DSBs, direct tests of this hypothesis are lacking. Here, we developed an in vivo single DSB system for both heterochromatic and euchromatic loci in Drosophila melanogaster Live imaging of single DSBs in larval imaginal discs recapitulates the spatio-temporal dynamics observed for irradiation (IR)-induced breaks in cell culture. Importantly, live imaging and sequence analysis of repair products reveal that DSBs in euchromatin and heterochromatin are repaired with similar kinetics, employ both NHEJ and HR, and can use homologous chromosomes as an HR template. This direct analysis reveals important insights into heterochromatin DSB repair in animal tissues and provides a foundation for further explorations of repair mechanisms in different chromatin domains.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
National Institutes of Health (NIH); USDOE
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1379517
Alternate ID(s):
OSTI ID: 1379518
Journal Information:
Genes & Development, Vol. 30, Issue 14; ISSN 0890-9369
Publisher:
Cold Springs Harbor PressCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 69 works
Citation information provided by
Web of Science

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Cited By (28)

A limited number of double-strand DNA breaks is sufficient to delay cell cycle progression journal September 2018
Timely double-strand break repair and pathway choice in pericentromeric heterochromatin depend on the histone demethylase dKDM4A journal December 2018
A limited number of double-strand DNA breaks are sufficient to delay cell cycle progression journal May 2018
Chromatin and nucleosome dynamics in DNA damage and repair journal November 2017
Recombination at subtelomeres is regulated by physical distance, double‐strand break resection and chromatin status journal July 2017
The response to DNA damage in heterochromatin domains journal March 2018
Keep moving and stay in a good shape to find your homologous recombination partner journal August 2018
Chromosome Preference During Homologous Recombination Repair of DNA Double-Strand Breaks in Drosophila melanogaster journal September 2019
BLM’s balancing act and the involvement of FANCJ in DNA repair journal September 2018
Quantification of DNA damage induced γH2AX focus formation via super-resolution dSTORM localization microscopy posted_content May 2019
And yet, it moves: nuclear and chromatin dynamics of a heterochromatic double-strand break
  • Caridi, P. Christopher; Delabaere, Laetitia; Zapotoczny, Grzegorz
  • Philosophical Transactions of the Royal Society B: Biological Sciences, Vol. 372, Issue 1731 https://doi.org/10.1098/rstb.2016.0291
journal August 2017
Timely double-strand break repair and pathway choice in pericentromeric heterochromatin depend on the histone demethylase dKDM4A journal April 2018
Heterochromatin delays CRISPR-Cas9 mutagenesis but does not influence repair outcome posted_content February 2018
The Importance of Satellite Sequence Repression for Genome Stability journal January 2017
Arp2/3 and Unc45 maintain heterochromatin stability in Drosophila polytene chromosomes journal July 2019
DNA end-resection in highly accessible chromatin produces a toxic break posted_content August 2021
Epigenetics of skin cancer: Interventions by selected bioactive phytochemicals
  • Penta, Dhanamjai; Somashekar, Bagganahalli S.; Meeran, Syed Musthapa
  • Photodermatology, Photoimmunology & Photomedicine, Vol. 34, Issue 1 https://doi.org/10.1111/phpp.12353
journal November 2017
DNA Repair in Drosophila : Mutagens, Models, and Missing Genes journal February 2017
A versatile genetic tool for post-translational control of gene expression in Drosophila melanogaster journal November 2017
Nuclear F-actin and myosins drive relocalization of heterochromatic breaks journal June 2018
Quantification of DNA damage induced repair focus formation via super-resolution dSTORM localization microscopy journal January 2019
Ten principles of heterochromatin formation and function journal December 2017
Jumonji Inhibitors Overcome Radioresistance in Cancer through Changes in H3K4 Methylation at Double-Strand Breaks journal October 2018
Chromatin Dynamics in Genome Stability: Roles in Suppressing Endogenous DNA Damage and Facilitating DNA Repair journal July 2017
Cervantes and Quijote protect heterochromatin from aberrant recombination and lead the way to the nuclear periphery journal September 2016
Arp2/3 and Unc45 maintain heterochromatin stability in Drosophila polytene chromosomes journal June 2019
Sleep increases chromosome dynamics to enable reduction of accumulating DNA damage in single neurons journal March 2019
Nuclear actin filaments in DNA repair dynamics journal September 2019

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