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

Yi Zhou | Zendy; Pierre Caron | Zendy; Gaëlle Legube | Zendy; Tanya T. Paull | Zendy

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

Author(s) -

Yi Zhou,

Pierre Caron,

Gaëlle Legube,

Tanya T. Paull

Publication year - 2013

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkt1309

Subject(s) - rad51 , homologous recombination , dna , genome instability , dna repair , dna damage , endonuclease , microbiology and biotechnology , chemistry , protein subunit , dna clamp , biology , biochemistry , gene , polymerase chain reaction , reverse transcriptase

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.

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