CtIP-mediated resection is essential for viability and can operate independently of BRCA1

Federica Polato, Elsa Callen, Nancy Wong, Robert Faryabi, Samuel Bunting, Hua Tang Chen, Marina Kozak, Michael J. Kruhlak, Colleen R. Reczek, Wen Hwa Lee, Thomas Ludwig, Richard Baer, Lionel Feigenbaum, Stephen Jackson, André Nussenzweig*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

69 Scopus citations

Abstract

Homologous recombination (HR) is initiated by DNA end resection, a process in which stretches of single-strand DNA (ssDNA) are generated and used for homology search. Factors implicated in resection include nucleases MRE11, EXO1, and DNA2, which process DNA ends into 39 ssDNA overhangs; helicases such as BLM, which unwind DNA; and other proteins such as BRCA1 and CtIP whose functions remain unclear. CDK-mediated phosphorylation of CtIP on T847 is required to promote resection, whereas CDK-dependent phosphorylation of CtIP-S327 is required for interaction with BRCA1. Here, we provide evidence that CtIP functions independently of BRCA1 in promoting DSB end resection. First, using mouse models expressing S327A or T847A mutant CtIP as a sole species, and B cells deficient in CtIP, we show that loss of the CtIP-BRCA1 interaction does not detectably affect resection, maintenance of genomic stability or viability, whereas T847 is essential for these functions. Second, although loss of 53BP1 rescues the embryonic lethality and HR defects in BRCA1-deficient mice, it does not restore viability or genome integrity in CtIP-/- mice. Third, the increased resection afforded by loss of 53BP1 and the rescue of BRCA1-deficiency depend on CtIP but not EXO1. Finally, the sensitivity of BRCA1-deficient cells to poly ADP ribose polymerase (PARP) inhibition is partially rescued by the phospho-mimicking mutant CtIP (CtIP-T847E). Thus, in contrast to BRCA1, CtIP has indispensable roles in promoting resection and embryonic development.

Original languageEnglish (US)
Pages (from-to)1027-1036
Number of pages10
JournalJournal of Experimental Medicine
Volume211
Issue number6
DOIs
StatePublished - Jun 2014

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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