DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination

Ronja Biehs; Monika Steinlage; Olivia Barton; Szilvia Juhász; Julia Künzel; Julian Spies; Atsushi Shibata; Penny A Jeggo; Markus Löbrich

doi:10.1016/j.molcel.2016.12.016

DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination

Mol Cell. 2017 Feb 16;65(4):671-684.e5. doi: 10.1016/j.molcel.2016.12.016. Epub 2017 Jan 26.

Authors

Ronja Biehs¹, Monika Steinlage¹, Olivia Barton¹, Szilvia Juhász¹, Julia Künzel¹, Julian Spies¹, Atsushi Shibata², Penny A Jeggo³, Markus Löbrich⁴

Affiliations

¹ Radiation Biology and DNA Repair, Darmstadt University of Technology, 64287 Darmstadt, Germany.
² Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, UK; Advanced Scientific Research Leaders Development Unit, Gunma University, Maebashi, Gunma 371-8511, Japan. Electronic address: shibata.at@gunma-u.ac.jp.
³ Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, UK. Electronic address: p.a.jeggo@sussex.ac.uk.
⁴ Radiation Biology and DNA Repair, Darmstadt University of Technology, 64287 Darmstadt, Germany. Electronic address: lobrich@bio.tu-darmstadt.de.

Abstract

Canonical non-homologous end joining (c-NHEJ) repairs DNA double-strand breaks (DSBs) in G1 cells with biphasic kinetics. We show that DSBs repaired with slow kinetics, including those localizing to heterochromatic regions or harboring additional lesions at the DSB site, undergo resection prior to repair by c-NHEJ and not alt-NHEJ. Resection-dependent c-NHEJ represents an inducible process during which Plk3 phosphorylates CtIP, mediating its interaction with Brca1 and promoting the initiation of resection. Mre11 exonuclease, EXD2, and Exo1 execute resection, and Artemis endonuclease functions to complete the process. If resection does not commence, then repair can ensue by c-NHEJ, but when executed, Artemis is essential to complete resection-dependent c-NHEJ. Additionally, Mre11 endonuclease activity is dispensable for resection in G1. Thus, resection in G1 differs from the process in G2 that leads to homologous recombination. Resection-dependent c-NHEJ significantly contributes to the formation of deletions and translocations in G1, which represent important initiating events in carcinogenesis.

Keywords: DNA double-strand breaks; non-homologous end joining; nucleases; resection.

Publication types

Comparative Study

MeSH terms

BRCA1 Protein / genetics
BRCA1 Protein / metabolism
Carrier Proteins / genetics
Carrier Proteins / metabolism
Cell Nucleus / enzymology
Cell Nucleus / pathology
Cell Nucleus / radiation effects*
DNA Breaks, Double-Stranded*
DNA End-Joining Repair / radiation effects*
DNA Repair Enzymes / genetics
DNA Repair Enzymes / metabolism
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Endodeoxyribonucleases
Endonucleases
Exodeoxyribonucleases / genetics
Exodeoxyribonucleases / metabolism
G1 Phase / radiation effects*
G2 Phase
Gene Deletion
HeLa Cells
Humans
Kinetics
MRE11 Homologue Protein
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Phosphorylation
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
Time Factors
Transfection
Translocation, Genetic
Tumor Suppressor Proteins
Tumor Suppressor p53-Binding Protein 1 / genetics
Tumor Suppressor p53-Binding Protein 1 / metabolism

Substances

BRCA1 Protein
BRCA1 protein, human
Carrier Proteins
DNA-Binding Proteins
MRE11 protein, human
Nuclear Proteins
TP53BP1 protein, human
Tumor Suppressor Proteins
Tumor Suppressor p53-Binding Protein 1
PLK3 protein, human
Protein Serine-Threonine Kinases
DCLRE1C protein, human
EXO1 protein, human
Endodeoxyribonucleases
Endonucleases
Exodeoxyribonucleases
MRE11 Homologue Protein
RBBP8 protein, human
DNA Repair Enzymes

Abstract

Publication types

MeSH terms

Substances

Grants and funding