Quantitation of DNA double-strand break resection intermediates in human cells.

Abstract:

:5' strand resection at DNA double strand breaks (DSBs) is critical for homologous recombination (HR) and genomic stability. Here we develop a novel method to quantitatively measure single-stranded DNA intermediates in human cells and find that the 5' strand at endonuclease-generated break sites is resected up to 3.5 kb in a cell cycle-dependent manner. Depletion of CtIP, Mre11, Exo1 or SOSS1 blocks resection, while depletion of 53BP1, Ku or DNA-dependent protein kinase catalytic subunit leads to increased resection as measured by this method. While 53BP1 negatively regulates DNA end processing, depletion of Brca1 does not, suggesting that the role of Brca1 in HR is primarily to promote Rad51 filament formation, not to regulate end resection.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Zhou Y,Caron P,Legube G,Paull TT

doi

10.1093/nar/gkt1309

subject

Has Abstract

pub_date

2014-02-01 00:00:00

pages

e19

issue

3

eissn

0305-1048

issn

1362-4962

pii

gkt1309

journal_volume

42

pub_type

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