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Structural basis for DNA 5´-end resection by RecJ.

Kaiying Cheng1, Hong Xu1, Xuanyi Chen1

  • 1Key Laboratory of Chinese Ministry of Agriculture for Nuclear-Agricultural Sciences, Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou, China.

Elife
|April 9, 2016
PubMed
Summary

RecJ protein is crucial for DNA repair by resecting DNA at double-strand breaks. Structural insights reveal how RecJ and SSB protein work together to efficiently process broken DNA ends for homologous recombination.

Keywords:
DNA end resectionDeinococcus radioduransRecF pathwayRecJbiophysicssingle-strand-DNA binding proteinstructural biologytwo-metal-ion catalysis

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • DNA double-strand breaks trigger recombinational repair pathways.
  • RecJ is a key 5´ nuclease in the RecF recombination pathway, essential for DNA end processing.

Purpose of the Study:

  • To elucidate the structural basis of RecJ's function in DNA repair.
  • To understand the mechanism of RecJ-mediated DNA end resection and its interaction with SSB.

Main Methods:

  • X-ray crystallography was used to determine the structures of Deinococcus radiodurans RecJ.
  • Complex structures were solved with deoxythymidine monophosphate (dTMP), single-stranded DNA (ssDNA), and the C-terminal region of single-stranded DNA-binding protein (SSB-Ct).

Main Results:

  • A 5´-phosphate-binding pocket dictates RecJ's 5´-3´ exonuclease polarity.
  • A helical gateway selectively admits ssDNA, and extensive protein-nucleotide interactions ensure processive resection.
  • The active site contains two divalent cations coordinating nucleophilic water, and ssDNA undergoes a 180° turn.
  • RecJ's C-terminal domain binds SSB-Ct, facilitating collaborative DNA processing.

Conclusions:

  • The determined structures provide mechanistic insights into the RecF pathway.
  • RecJ's unique structural features enable efficient and processive DNA end resection.
  • The interaction between RecJ and SSB is critical for homologous recombination repair.