An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing

Hideki Aihara, Yutaka Ito, Hitoshi Kurumizaka, Tohru Terada, Shigeyuki Yokoyama, Takehiko Shibata

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

RecA protein and its homologs catalyze homologous pairing of dsDNA and ssDNA, a critical reaction in homologous genetic recombination in various organisms from a virus, microbes to higher eukaryotes. In this reaction, RecA protein forms a nucleoprotein filament on ssDNA, which in turn binds to naked dsDNA for homology search. We suggested that the C-terminal domain of RecA protein plays a role in capturing the dsDNA. Here, we isolated the C-terminal domain as a soluble form and determined the solution structure by NMR spectroscopy. The overall folding of the NMR structure agrees with that of the corresponding part of the reported crystal structure, but a remarkable difference was found in a solvent-exposed region due to intermolecular contacts in the crystal. Then, we studied the interaction between the C-terminal domain and DNA, and found that significant chemical shift changes were induced in a specific region by titration with dsDNA. SsDNA induced a much smaller chemical shift perturbation. The difference of DNA concentrations to give the half-saturation of the chemical shift change showed a higher affinity of the C-terminal region toward dsDNA. Combined with our previous results, these provide direct evidence that the defined region in the C-terminal domain furnishes a binding surface for DNA.

Original languageEnglish
Pages (from-to)213-221
Number of pages9
JournalJournal of Molecular Biology
Volume274
Issue number2
DOIs
Publication statusPublished - 1997 Nov 28
Externally publishedYes

Fingerprint

Rec A Recombinases
DNA
Nucleoproteins
Homologous Recombination
Eukaryota
Genetic Recombination
Magnetic Resonance Spectroscopy
Viruses

Keywords

  • DNA-binding surface
  • Genetic recombination
  • NMR
  • Solution structure
  • X-ray structure

ASJC Scopus subject areas

  • Virology

Cite this

An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing. / Aihara, Hideki; Ito, Yutaka; Kurumizaka, Hitoshi; Terada, Tohru; Yokoyama, Shigeyuki; Shibata, Takehiko.

In: Journal of Molecular Biology, Vol. 274, No. 2, 28.11.1997, p. 213-221.

Research output: Contribution to journalArticle

Aihara, Hideki ; Ito, Yutaka ; Kurumizaka, Hitoshi ; Terada, Tohru ; Yokoyama, Shigeyuki ; Shibata, Takehiko. / An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing. In: Journal of Molecular Biology. 1997 ; Vol. 274, No. 2. pp. 213-221.
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