CtIP protein dimerization is critical for its recruitment to chromosomal DNA double-stranded breaks

Hailong Wang, Zhengping Shao, Linda Z. Shi, Patty Yi Hwa Hwang, Lan N. Truong, Michael W. Berns, David J. Chen, Xiaohua Wu

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46 Scopus citations

Abstract

CtIP (CtBP-interacting protein) associates with BRCA1 and the Mre11-Rad50-Nbs1 (MRN) complex and plays an essential role in homologous recombination (HR)-mediated DNA double-stranded break (DSB) repair. It has been described that CtIP forms dimers in mammalian cells, but the biological significance is not clear. In this study, we identified a conserved motif in the N terminus of CtIP, which is required for dimer formation. We further showed that CtIP mutants impaired in forming dimers are strongly defective in HR, end resection, and activation of the ataxia telangiectasia and Rad3-related pathway, without notable change of CtIP interactions with BRCA1 or Nbs1. In addition to HR, CtIP dimerization is also required for microhomology-mediated end joining. Live cell imaging of enhanced GFP-tagged CtIP demonstrates that the CtIP dimerization mutant fails to be localized to DSBs, whereas placing a heterologous dimerization motif to the dimerization mutant restores CtIP recruitment to DSBs. These studies suggest that CtIP dimer formation is essential for its recruitment to DSBs on chromatin upon DNA damage. Furthermore, DNA damage-induced phosphorylation of CtIP is significantly reduced in the CtIP dimerization mutants. Therefore, in addition to the C-terminal conserved domains critical for CtIP function, the dimerization motif on the N terminus of CtIP is also conserved and essential for its function in DNA damage responses. The severe repair defects of CtIP dimerization mutants are likely due to the failure in localization to chromosomal DSBs upon DNA damage.

Original languageEnglish (US)
Pages (from-to)21471-21480
Number of pages10
JournalJournal of Biological Chemistry
Volume287
Issue number25
DOIs
StatePublished - Jun 15 2012

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wang, H., Shao, Z., Shi, L. Z., Hwang, P. Y. H., Truong, L. N., Berns, M. W., Chen, D. J., & Wu, X. (2012). CtIP protein dimerization is critical for its recruitment to chromosomal DNA double-stranded breaks. Journal of Biological Chemistry, 287(25), 21471-21480. https://doi.org/10.1074/jbc.M112.355354