Tetrameric Ctp1 coordinates DNA binding and DNA bridging in DNA double-strand-break repair

Nat Struct Mol Biol. 2015 Feb;22(2):158-66. doi: 10.1038/nsmb.2945. Epub 2015 Jan 12.

Abstract

Ctp1 (also known as CtIP or Sae2) collaborates with Mre11-Rad50-Nbs1 to initiate repair of DNA double-strand breaks (DSBs), but its functions remain enigmatic. We report that tetrameric Schizosaccharomyces pombe Ctp1 contains multivalent DNA-binding and DNA-bridging activities. Through structural and biophysical analyses of the Ctp1 tetramer, we define the salient features of Ctp1 architecture: an N-terminal interlocking tetrameric helical dimer-of-dimers (THDD) domain and a central intrinsically disordered region (IDR) linked to C-terminal 'RHR' DNA-interaction motifs. The THDD, IDR and RHR are required for Ctp1 DNA-bridging activity in vitro, and both the THDD and RHR are required for efficient DSB repair in S. pombe. Our results establish non-nucleolytic roles of Ctp1 in binding and coordination of DSB-repair intermediates and suggest that ablation of human CtIP DNA binding by truncating mutations underlie the CtIP-linked Seckel and Jawad syndromes.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA Breaks, Double-Stranded
  • DNA Repair / physiology
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism*
  • Protein Binding
  • Protein Multimerization / physiology*
  • Schizosaccharomyces
  • Schizosaccharomyces pombe Proteins / chemistry*
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Ctp1 protein, S pombe
  • DNA-Binding Proteins
  • Schizosaccharomyces pombe Proteins

Associated data

  • PDB/4X01